├── .gitignore
├── .gitmodules
├── LICENSE
├── README.md
├── all.fa.gz
├── bin
├── Linux_x86_64_kernel_3.10.0
│ └── quack
├── MacOS_10.14.6
│ └── quack
└── Windows_Cygwin_2.904_x86_64
│ └── quack.exe
├── images
├── .gitignore
├── explanation.overlay.svg
├── explanation.png
├── makefile
├── paired.adapter.png
├── paired.adapter.svg
├── paired.noadapt.png
├── paired.noadapt.svg
├── single.adapter.png
├── single.adapter.svg
├── single.noadapt.png
└── single.noadapt.svg
├── makefile
├── quack.c
├── svg.c
└── svg.h
/.gitignore:
--------------------------------------------------------------------------------
1 | *~
2 | *.o
3 | quack
4 |
--------------------------------------------------------------------------------
/.gitmodules:
--------------------------------------------------------------------------------
1 | [submodule "klib"]
2 | path = klib
3 | url = https://github.com/attractivechaos/klib
4 |
--------------------------------------------------------------------------------
/LICENSE:
--------------------------------------------------------------------------------
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570 | Each version is given a distinguishing version number. If the
571 | Program specifies that a certain numbered version of the GNU General
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573 | option of following the terms and conditions either of that numbered
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575 | Foundation. If the Program does not specify a version number of the
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578 |
579 | If the Program specifies that a proxy can decide which future
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587 | later version.
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589 | 15. Disclaimer of Warranty.
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592 | APPLICABLE LAW. EXCEPT WHEN OTHERWISE STATED IN WRITING THE COPYRIGHT
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621 | END OF TERMS AND CONDITIONS
622 |
623 | How to Apply These Terms to Your New Programs
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625 | If you develop a new program, and you want it to be of the greatest
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652 | If the program does terminal interaction, make it output a short
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667 | .
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669 | The GNU General Public License does not permit incorporating your program
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671 | may consider it more useful to permit linking proprietary applications with
672 | the library. If this is what you want to do, use the GNU Lesser General
673 | Public License instead of this License. But first, please read
674 | .
675 |
--------------------------------------------------------------------------------
/README.md:
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1 | # quack
2 |
3 | A FASTQ quality assessment tool
4 |
5 | ## Citation
6 | A. Thrash, M. Arick, and D. G. Peterson, “Quack: A quality assurance tool for high throughput sequence data,” Analytical Biochemistry, vol. 548, pp. 38–43, 2018. https://doi.org/10.1016/j.ab.2018.01.028
7 |
8 | ## Latest Release
9 |
10 | The latest release of quack and its binaries can always be found [here](https://github.com/IGBB/quack/releases/latest).
11 |
12 | ## Dependencies
13 |
14 | * zlib
15 | * klib (pulled by the submodule update below)
16 |
17 | ## Installation from Source
18 |
19 | git clone https://github.com/IGBB/quack.git
20 |
21 | cd quack/
22 |
23 | make && make test
24 |
25 | ## Binaries
26 |
27 | Binaries are available in the bin/ folder. Current testing of these binaries has been limited. If a binary doesn't work, try compiling from source on your system.
28 |
29 | ## Running Quack
30 |
31 | Quack has the following options.
32 |
33 | ```
34 | -1, --forward forward strand data in gzipped FASTQ format, must be used with -2 or --reverse
35 | -2, --reverse reverse strand data in gzipped FASTQ format, must be used with -1 or --forward
36 | -a, --adapters adapters in gzipped FASTA format (optional)
37 | -n, --name a descriptive name to be printed with the output image (optional)
38 | -u, --unpaired unpaired data in gzipped FASTQ format
39 | -?, --help, --usage prints the help or usage information
40 | -V, --version prints the program version
41 | ```
42 |
43 | Quack takes gzipped FASTQ-formatted files as input for data and gzipped As output, quack prints an SVG formatted image to standard output.
44 |
45 |
46 | ### Examples
47 |
48 | #### Paired-end with name and adapters
49 | `quack -1 reads.1.fastq.gz -2 reads.2.fastq.gz -n sample_name -a adapters_files.fasta.gz > sample_name.svg`
50 |
51 | #### Unpaired with name and adapters
52 | `quack -u reads.fastq.gz -n sample_name -a adapters.fa.gz > sample_name.svg`
53 |
54 | #### Paired-end without name and adapters
55 | `quack -1 reads.1.fastq.gz -2 reads.2.fastq.gz > sample_name.svg`
56 |
57 | #### Unpaired without name and adapters
58 | `quack -u reads.fastq.gz > sample_name.svg`
59 |
60 | ### Output
61 |
62 | Quack is capable of producing output for single-ended data and paired-end data. Only the singled-ended data is labeled, since the paried-end data has all the same parts.
63 |
64 | #### Single-ended Data
65 | 
66 |
67 |
68 | A. The base content distribution showing the percentage of each nucleotide in each column of an array.
69 | B. A heatmap showing the distribution of sequence quality for each column and a line representing mean quality scores across the array
70 | C. A score distribution graph showing the percentage of bases matching certain scores, with 100% on the left of the graph and 0% on the right. The highest scoring data appears at the top of the graph.
71 | D. Length distribution graph showing the percentage of reads of a given length
72 | E. Adapter content distribution graph showing how adapter content is distributed throughout an array
73 |
74 | #### Paired-end Data
75 | 
76 |
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/all.fa.gz:
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https://raw.githubusercontent.com/IGBB/quack/cf394194d22884e26ba470c0442aaf83a0f8a842/all.fa.gz
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/bin/Linux_x86_64_kernel_3.10.0/quack:
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https://raw.githubusercontent.com/IGBB/quack/cf394194d22884e26ba470c0442aaf83a0f8a842/bin/Linux_x86_64_kernel_3.10.0/quack
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/bin/MacOS_10.14.6/quack:
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https://raw.githubusercontent.com/IGBB/quack/cf394194d22884e26ba470c0442aaf83a0f8a842/bin/MacOS_10.14.6/quack
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/bin/Windows_Cygwin_2.904_x86_64/quack.exe:
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https://raw.githubusercontent.com/IGBB/quack/cf394194d22884e26ba470c0442aaf83a0f8a842/bin/Windows_Cygwin_2.904_x86_64/quack.exe
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/images/.gitignore:
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1 | SRR1168757.fastq.gz
2 | ERR1438847_1.fastq.gz
3 | ERR1438847_2.fastq.gz
4 |
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/images/explanation.overlay.svg:
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1 |
13 |
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/images/explanation.png:
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https://raw.githubusercontent.com/IGBB/quack/cf394194d22884e26ba470c0442aaf83a0f8a842/images/explanation.png
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/images/makefile:
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1 |
2 | SRR1168757.fastq.gz:
3 | wget ftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR116/007/SRR1168757/$@
4 |
5 | ERR1438847_%.fastq.gz:
6 | wget ftp://ftp.sra.ebi.ac.uk/vol1/fastq/ERR143/007/ERR1438847/$@
7 |
8 | single.adapter.svg : ../quack SRR1168757.fastq.gz
9 | ../quack -u SRR1168757.fastq.gz -a ../all.fa.gz -n SRR1168757 > $@
10 |
11 | single.noadapt.svg : ../quack SRR1168757.fastq.gz
12 | ../quack -u SRR1168757.fastq.gz -n SRR1168757 > $@
13 |
14 | paired.adapter.svg : ../quack ERR1438847_1.fastq.gz ERR1438847_2.fastq.gz
15 | ../quack -1 ERR1438847_1.fastq.gz -2 ERR1438847_2.fastq.gz -a ../all.fa.gz -n ERR1438847 > $@
16 |
17 | paired.noadapt.svg : ../quack ERR1438847_1.fastq.gz ERR1438847_2.fastq.gz
18 | ../quack -1 ERR1438847_1.fastq.gz -2 ERR1438847_2.fastq.gz -n ERR1438847 > $@
19 |
20 | explanation.overlay.png : explanation.overlay.svg
21 | convert -background none $< $@
22 |
23 | explanation.png : explanation.overlay.png single.adapter.png
24 | composite -gravity center $^ $@
25 |
26 | %.png: %.svg
27 | convert $< $@
28 |
29 | .PHONY: clean all
30 | clean:
31 | rm -f $(obj) quack ERR1438847_1.fastq.gz ERR1438847_2.fastq.gz
32 |
33 | all: paired.adapter.png \
34 | paired.noadapt.png \
35 | single.adapter.png \
36 | single.noadapt.png \
37 | explanation.png
38 |
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/images/paired.noadapt.png:
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https://raw.githubusercontent.com/IGBB/quack/cf394194d22884e26ba470c0442aaf83a0f8a842/images/paired.noadapt.png
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/images/single.adapter.png:
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https://raw.githubusercontent.com/IGBB/quack/cf394194d22884e26ba470c0442aaf83a0f8a842/images/single.adapter.png
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/images/single.noadapt.png:
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https://raw.githubusercontent.com/IGBB/quack/cf394194d22884e26ba470c0442aaf83a0f8a842/images/single.noadapt.png
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/images/single.noadapt.svg:
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1 |
679 |
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/makefile:
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1 | src = $(wildcard *.c)
2 | obj = $(src:.c=.o)
3 |
4 | override LDFLAGS := -lz -lm $(LDFLAGS)
5 | override CFLAGS := -Iklib -O3 $(CFLAGS)
6 |
7 | all : klib/kseq.h quack
8 |
9 | quack: $(obj)
10 | $(CC) -o $@ $^ $(LDFLAGS)
11 |
12 | klib/kseq.h:
13 | git submodule update --init --recursive
14 |
15 | %.o: %.c
16 | $(CC) $(CFLAGS) -o $@ -c $<
17 |
18 | .PHONY: all clean images test
19 | clean:
20 | rm -f $(obj) quack
21 |
22 | images: quack
23 | $(MAKE) -C images all
24 |
25 | test: images
26 |
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/quack.c:
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1 | #include
2 | #include
3 | #include
4 | #include
5 | #include
6 | #include
7 |
8 | #include "kseq.h"
9 | #include "svg.h"
10 |
11 | #define unlikely(x) __builtin_expect ((x), 0)
12 | #define likely(x) __builtin_expect((x),1)
13 |
14 |
15 | #define svg_axis_label( posx, posy, rot, label) \
16 | svg_start_tag("text", 7, \
17 | svg_attr("x", "%d", posx), \
18 | svg_attr("fill", "%s", "#AAA"), \
19 | svg_attr("y", "%d", posy), \
20 | svg_attr("font-family", "%s", "sans-serif"), \
21 | svg_attr("font-size", "%s", "15px"), \
22 | svg_attr("text-anchor", "%s", "middle"), \
23 | svg_attr("transform", "rotate(%d)", rot) \
24 | ); \
25 | printf("%s\n", label); \
26 | svg_end_tag("text");
27 | #define svg_axis_number( posx, posy, a, number) \
28 | svg_start_tag("text", 6, \
29 | svg_attr("x", "%d", posx), \
30 | svg_attr("fill", "%s", "#AAA"), \
31 | svg_attr("y", "%d", posy), \
32 | svg_attr("font-family", "%s", "sans-serif"), \
33 | svg_attr("font-size", "%s", "10px"), \
34 | svg_attr("text-anchor", "%s", a) \
35 | ); \
36 | printf("%d\n", number); \
37 | svg_end_tag("text");
38 |
39 | #define svg_center_label( posx, posy, fillv, label_format, label) \
40 | svg_start_tag("text", 7, \
41 | svg_attr("x", "%d", posx), \
42 | svg_attr("y", "%d", posy), \
43 | svg_attr("fill", "%s", fillv), \
44 | svg_attr("font-family", "%s", "sans-serif"), \
45 | svg_attr("font-size", "%s", "15px"), \
46 | svg_attr("font-weight", "%s", "bold"), \
47 | svg_attr("text-anchor", "%s", "middle") \
48 | ); \
49 | printf(label_format, label); \
50 | svg_end_tag("text");
51 |
52 |
53 |
54 | const char *program_version = "quack 1.1.1";
55 | struct arguments {
56 | char *name, *forward, *reverse, *unpaired, *adapters;
57 | };
58 |
59 | struct arguments parse_options(int argc, char **argv) {
60 | struct arguments arguments = {
61 | .unpaired = NULL,
62 | .forward = NULL,
63 | .reverse = NULL,
64 | .name = NULL,
65 | .adapters = NULL
66 | };
67 |
68 | if (argc== 1 || argc == 2) {
69 | if (argc == 1 || (strcmp(argv[1], "--help") == 0 || strcmp(argv[1], "--usage") == 0 || strcmp(argv[1], "-?") == 0)) {
70 | printf("Usage: quack [OPTION...]\n"
71 | "quack -- A FASTQ quality assessment tool\n\n"
72 | " -1, --forward file.1.fq.gz Forward strand\n"
73 | " -2, --reverse file.2.fq.gz Reverse strand\n"
74 | " -a, --adapters adapters.fa.gz (Optional) Adapters file\n"
75 | " -n, --name NAME (Optional) Display in output\n"
76 | " -u, --unpaired unpaired.fq.gz Data (only use with -u)\n"
77 | " -?, --help Give this help list\n"
78 | " --usage (use alone)\n"
79 | " -V, --version Print program version (use alone)\n"
80 | "Report bugs to .\n");
81 | }
82 |
83 | if (argc == 2 && ((strcmp(argv[1], "-V") == 0 || strcmp(argv[1], "--version") == 0))) {
84 | printf("%s\n", program_version);
85 | exit (0);
86 | }
87 | }
88 |
89 | if(argc>2 && argc % 2 != 0)
90 | {
91 | int counter=1;
92 | while (counter.\n");
124 |
125 | exit(EXIT_FAILURE);
126 | }
127 |
128 | counter = counter+2;
129 | }
130 | }
131 | return arguments;
132 | }
133 |
134 | typedef struct {
135 | uint64_t scores[91];
136 | uint64_t content[4];
137 | uint64_t length_count;
138 | uint64_t kmer_count;
139 | } base_information;
140 |
141 | typedef struct {
142 | base_information *bases;
143 | uint64_t max_length;
144 | uint64_t original_max_length;
145 | uint64_t number_of_sequences;
146 | } sequence_data;
147 |
148 | /* Convert ASCII to Integer for A T C and G */
149 | /* A C G T */
150 | int lookup[20] = {0, 0, 2, 0, 0, 0, 3, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 0, 1};
151 |
152 | KSEQ_INIT(gzFile, gzread)
153 |
154 | int* read_adapters(char *adapters_file) {
155 | int kmer_size = 10;
156 | int array_size = pow(4, kmer_size);
157 | gzFile fp;
158 | kseq_t *seq;
159 | int i, l, index;
160 | fp = gzopen(adapters_file, "r");
161 | seq = kseq_init(fp);
162 | int *kmers = malloc(array_size*sizeof(int));
163 | memset(kmers, 0, array_size*sizeof(int));
164 | while ((l = kseq_read(seq)) >= 0) {
165 | index = 0;
166 | for (i = 0; i < kmer_size; i++) {
167 | index = ((index << 2) + (lookup[seq->seq.s[i]-65 & ~32])) & (array_size-1);
168 | }
169 | for (; i < seq->seq.l; i++) {
170 | index = ((index << 2) + (lookup[seq->seq.s[i]-65 & ~32])) & (array_size-1);
171 | kmers[index] = 1;
172 | }
173 |
174 | }
175 | kseq_destroy(seq);
176 | gzclose(fp);
177 | return kmers;
178 | }
179 |
180 | sequence_data* read_fastq(char *fastq_file, int *kmers) {
181 | gzFile fp;
182 | kseq_t *seq;
183 | int i, l, index;
184 | int kmer_size = 10;
185 | int array_size = pow(4, kmer_size);
186 | int max_length = 0;
187 | fp = gzopen(fastq_file, "r");
188 | seq = kseq_init(fp);
189 | base_information *bases = NULL;
190 | sequence_data *to_return = malloc(sizeof(sequence_data));
191 | int number_of_sequences = 0;
192 |
193 | while ((l = kseq_read(seq)) >= 0) {
194 | if (unlikely(seq->seq.l > max_length)) {
195 | bases = realloc(bases, seq->seq.l*sizeof(base_information));
196 | memset(bases+max_length, 0, (seq->seq.l - max_length)*sizeof(base_information));
197 | max_length = seq->seq.l;
198 | }
199 | for (i = 0; i < seq->seq.l; i++) {
200 | int base = seq->seq.s[i];
201 | int offset = lookup[base-65 & ~32];
202 | bases[i].content[offset]++;
203 | int quality = seq->qual.s[i]-33;
204 | bases[i].scores[quality]++;
205 | }
206 | index = 0;
207 | for (i = 0; i < kmer_size; i++) {
208 | index = ((index << 2) + (lookup[seq->seq.s[i]-65 & ~32])) & (array_size-1);
209 | }
210 | if (kmers) {
211 | for (; kmers[index] == 0 && i < seq->seq.l; i++) {
212 | index = ((index << 2) + (lookup[seq->seq.s[i]-65 & ~32])) & (array_size-1);
213 | }
214 | }
215 | if (i < seq->seq.l) {
216 | bases[i].kmer_count++;
217 | }
218 |
219 | bases[seq->seq.l-1].length_count++;
220 | number_of_sequences++;
221 | }
222 | kseq_destroy(seq);
223 | gzclose(fp);
224 | to_return->bases = bases;
225 | to_return->max_length = max_length;
226 | to_return->number_of_sequences = number_of_sequences;
227 | return to_return;
228 | }
229 |
230 | sequence_data* transform(sequence_data* data) {
231 | int i, j;
232 | data->original_max_length = data->max_length;
233 | // binning
234 | if (data->max_length > 3000) {
235 | fprintf(stderr, "Binning...\n");
236 | int bin_size = 100;
237 | int unbinned;
238 | int binned = 0;
239 |
240 | for (unbinned = 1; unbinned < data->max_length; unbinned++) {
241 | if (unbinned%bin_size == 0){
242 | binned++;
243 | for (i = 0; i < 4; i++) {
244 | data->bases[binned].content[i] = 0;
245 | }
246 | for (i = 0; i < 91; i++) {
247 | data->bases[binned].scores[i] = 0;
248 | }
249 | data->bases[binned].length_count = 0;
250 | }
251 | for (i = 0; i < 4; i++) {
252 | data->bases[binned].content[i] = data->bases[binned].content[i] + data->bases[unbinned].content[i];
253 | }
254 | for (i = 0; i < 91; i++) {
255 | data->bases[binned].scores[i] = data->bases[binned].scores[i] + data->bases[unbinned].scores[i];
256 | }
257 | // fprintf(stderr, "%d\n", data->bases[binned].length_count);
258 | data->bases[binned].length_count = data->bases[binned].length_count + data->bases[unbinned].length_count;
259 | data->bases[binned].kmer_count = data->bases[binned].kmer_count + data->bases[unbinned].kmer_count;
260 | }
261 | data->max_length = binned;
262 | }
263 |
264 | for (i = 1; i < data->max_length; i++) {
265 | data->bases[i].kmer_count = data->bases[i-1].kmer_count + data->bases[i].kmer_count;
266 | }
267 |
268 | // transforming data for drawing (percentages rather than counts)
269 | for (i = 0; i < data->max_length; i++) {
270 | int content_sum = 0;
271 | int score_sum = 0;
272 | /* for (j = 0; j < 4; j++) { */
273 | /* content_sum = content_sum + data->bases[i].content[j]; */
274 | /* } */
275 | /* if (content_sum != 0) { */
276 | /* for (j = 0; j < 4; j++) { */
277 | /* data->bases[i].content[j] = 100*data->bases[i].content[j]/content_sum; */
278 | /* } */
279 | /* } */
280 | for (j = 0; j < 91; j++) {
281 | score_sum = score_sum + data->bases[i].scores[j];
282 | }
283 | if (score_sum != 0) {
284 | for (j = 0; j < 91; j++) {
285 | data->bases[i].scores[j] = 100*data->bases[i].scores[j]/score_sum;
286 | }
287 | }
288 | data->bases[i].length_count = ceil(100*(float)data->bases[i].length_count/data->number_of_sequences);
289 | data->bases[i].kmer_count = ceil(100*(float)data->bases[i].kmer_count/(float)data->number_of_sequences);
290 |
291 | }
292 | return data;
293 | }
294 |
295 | void draw(sequence_data* data, int position, int adapters_used) {
296 | int i, j, x, y;
297 | int counter = 0;
298 | int max_score = 0;
299 | uint64_t number_of_bases = 0;
300 | uint64_t total_counts[91] = {0};
301 | float averages[500];
302 |
303 | /********** Get Encoding ***************/
304 | char *encoding = "phred64";
305 | int offset = 31;
306 | /* Loop through every base */
307 | for( i=0; i < data->max_length; i++ ){
308 |
309 | /* Loop through scores, breaking when first non-zero is found for current
310 | * base */
311 | for(j=0; j < 31; j++){
312 | if(data->bases[i].scores[j] > 0) break;
313 | }
314 |
315 | /* Found encoding if j is lower than 31 (lower than possible for phred64
316 | * encoding) */
317 | if(j < 31){
318 | encoding="phred33";
319 | offset=0;
320 | break;
321 | }
322 | }
323 |
324 |
325 | // get max score, score distribution, and average scores at each position
326 | max_score = 40;
327 | for (i = 0; i < data->max_length; i++) {
328 | int sum = 0;
329 | for (j = offset; j < 91; j++) {
330 | if (data->bases[i].scores[j] > 0 && (j-offset) > max_score) {
331 | max_score = j - offset;
332 | }
333 | total_counts[j - offset] += data->bases[i].scores[j];
334 | sum += (j - offset)*data->bases[i].scores[j];
335 |
336 | }
337 | number_of_bases ++;
338 | /* score counts are already converted to percentage of total counts; so, all
339 | * that needs to be done to find average is divide by 100 */
340 | averages[i] = sum/100.0;
341 | }
342 |
343 |
344 | /********** File Stats ***************/
345 | svg_start_tag("text", 6,
346 | svg_attr("x", "%d", (position==0)?355:835),
347 | svg_attr("y", "%d", 20),
348 | svg_attr("text-anchor", "%s", "middle"),
349 | svg_attr("font-family", "%s", "sans-serif"),
350 | svg_attr("font-size", "%s", "15px"),
351 | svg_attr("fill", "%s", "#555")
352 | );
353 | svg_start_tag("tspan", 0);
354 | printf("%d", data->number_of_sequences);
355 | svg_end_tag("tspan");
356 | svg_start_tag("tspan", 1, svg_attr("fill", "%s", "#888"));
357 | printf(" reads with endcoding ");
358 | svg_end_tag("tspan");
359 | svg_start_tag("tspan", 0);
360 | printf("%s", encoding);
361 | svg_end_tag("tspan");
362 | svg_end_tag("text");
363 |
364 | /* Group for rug plot */
365 | svg_start_tag("g", 1,
366 | svg_attr("transform", "translate(%d %d)", 5, 25)
367 | );
368 |
369 | /* Horizontal Tick marks */
370 | x = 100;
371 | if (position==1)
372 | x = 1000;
373 | for (i = 10; i < 100; i+=10){
374 | y = 105 + i * 250 / 100;
375 |
376 | svg_simple_tag("line",6,
377 | svg_attr("x1", "%d", x),
378 | svg_attr("x2", "%d", x+100),
379 | svg_attr("y1", "%d", y),
380 | svg_attr("y2", "%d", y),
381 | svg_attr("stroke", "%s", "black"),
382 | svg_attr("stroke-width", "%f", (i%20 == 10)?1:0.5)
383 | );
384 | }
385 |
386 |
387 | /* Group for the vertical section of rug plot */
388 | svg_start_tag("g", 1,
389 | svg_attr("transform", "translate(%d 0)", (position == 1)?610:130)
390 | );
391 |
392 |
393 | /*************** Vertical Tick Marks ***************/
394 | y = (adapters_used == 0)?400:500;
395 | for (i = 10; i < 100; i+=10){
396 | x = i * 450 / 100;
397 | svg_simple_tag("line",6,
398 | svg_attr("x1", "%d", x),
399 | svg_attr("x2", "%d", x),
400 | svg_attr("y1", "%d", 10),
401 | svg_attr("y2", "%d", y),
402 | svg_attr("stroke", "%s", "black"),
403 | svg_attr("stroke-width", "%f", (i%20 == 10)?1:0.5)
404 | );
405 | }
406 |
407 |
408 | /*************** Base Ratio ***************/
409 |
410 | /* Flip svg to make svg coordinate system match cartesian coordinate. Must use
411 | a group since imagemagick uses SVG v1.1*/
412 | svg_start_tag("g", 1, svg_attr("transform", "translate(%d,%d) scale(%d, %d)", 0, 100, 1,-1));
413 |
414 | svg_start_tag("svg", 4,
415 | svg_attr("width", "%d", 450),
416 | svg_attr("height", "%d", 100),
417 | svg_attr("preserveAspectRatio", "%s", "none"),
418 | svg_attr("viewBox", "0 0 %d %d", data->max_length, data->number_of_sequences)
419 | );
420 |
421 | /* Set background color */
422 | svg_simple_tag("rect", 3,
423 | svg_attr("width", "%s", "100%"),
424 | svg_attr("height", "%s", "100%"),
425 | svg_attr("fill", "%s", "#CCC")
426 | );
427 |
428 |
429 | /* Allocate 25 characters per base (A,C,T,G) per point.
430 | 4 = max length is capped in kb range, will start compressing if larger
431 | 2 = '.5' added to point
432 | 1 = ','
433 | 15 = number of reads (10 quadrillion reads will break it)
434 | 1 = ' '
435 | 2 = padding for miscalculation
436 |
437 | */
438 | size_t ratio_points_length = 25*data->max_length;
439 | char * ratio_points[4];
440 | char tmp[25];
441 | for(i = 0; i < 4; i++){
442 | ratio_points[i] = malloc(ratio_points_length);
443 | }
444 |
445 | /* Since coordinates for lines and rectangles don't work the same; set the
446 | first point of each line to start off graph. Then, add 0.5 to the x of each
447 | point. Finally, end the line off graph. */
448 | y = 0;
449 | for(i = 0; i < 4; i++){
450 | y += data->bases[0].content[i];
451 | snprintf(ratio_points[i], ratio_points_length, "0,%d ", y);
452 | }
453 |
454 | /* Calculate cumlative sum for each x position and add it to point string */
455 | for (x = 0; x < data->max_length; x++) {
456 | y = 0;
457 | for(i = 0; i < 4; i++ ){
458 | y += data->bases[x].content[i];
459 |
460 | snprintf(tmp, 20, "%d.5,%d ", x, y);
461 | strncat(ratio_points[i], tmp, ratio_points_length);
462 | }
463 | }
464 |
465 |
466 | /* Make lines end off graph */
467 | y = 0;
468 | for(i = 0; i < 4; i++){
469 | y += data->bases[data->max_length-1].content[i];
470 |
471 | snprintf(tmp, 20, "%d,%d ", data->max_length, y);
472 | strncat(ratio_points[i], tmp, ratio_points_length);
473 | }
474 |
475 |
476 | /* Draw each distribution, in decending order so they stack */
477 | char *ratio_labels[4] = {"%A", "%T", "%C", "%G"};
478 | char *ratio_colors[4] = {"#648964", "#89bc89", "#84accf", "#5d7992"};
479 | for(i = 3; i >= 0; i--){
480 | svg_simple_tag("polyline", 3,
481 | svg_attr("points", "0,0 %s %d,0", ratio_points[i], data->max_length),
482 | svg_attr("fill", "%s", ratio_colors[i]),
483 | svg_attr("stroke", "%s", "none")
484 | );
485 | }
486 |
487 | for(i = 0; i < 4; i++)
488 | free(ratio_points[i]);
489 |
490 | svg_end_tag("svg"); // Base Ratio
491 | svg_end_tag("g"); // Base Ratio
492 |
493 | /* Lables */
494 |
495 | svg_start_tag("text", 5,
496 | svg_attr("y", "%d", 95),
497 | svg_attr("fill", "%s", "#CCC"),
498 | svg_attr("x", "%d", 5),
499 | svg_attr("font-family", "%s", "sans-serif"),
500 | svg_attr("font-size", "%s", "15px")
501 | );
502 | printf("%s\n", "Base Content Percentage");
503 | svg_end_tag("text");
504 |
505 |
506 | /* Print Ratio labels in center, only needed the first time
507 | 465 = width of vertical section (450) + width of margin (30) halved
508 | */
509 | if(position==0){
510 | for( i = 0; i < 4; i++){
511 | svg_center_label(465, 20*(4-i), ratio_colors[i], "%s", ratio_labels[i]);
512 | }
513 | }
514 |
515 | if(position == 0){
516 | svg_axis_label(-50, -5, -90, "Percent");
517 | svg_axis_number(-5, 100, "end", 0);
518 | svg_axis_number(-5, 5, "end", 100);
519 | }else{
520 | svg_axis_label(50, -455, 90, "Percent");
521 | svg_axis_number(455, 100, "start", 0);
522 | svg_axis_number(455, 5, "start", 100);
523 | }
524 |
525 | /*************** Heatmap ***************/
526 | /* Flip svg to make svg coordinate system match cartesian coordinate. The y
527 | is negative to compensate for the horizontal flip */
528 | svg_start_tag("g", 1, svg_attr("transform", "translate(%d,%d) scale(%d, %d)", 0, 355, 1,-1));
529 |
530 | svg_start_tag("svg", 4,
531 | svg_attr("width", "%d", 450),
532 | svg_attr("height", "%d", 250),
533 | svg_attr("preserveAspectRatio", "%s", "none"),
534 | svg_attr("viewBox", "0 0 %d %d", data->max_length, max_score)
535 | );
536 |
537 |
538 | /* Define background for heatmap. Must be in descending order or highest score
539 | background will overwrite all other backgrounds */
540 | #define score_back(score, color) \
541 | svg_simple_tag("rect", 6, \
542 | svg_attr("x", "%d", 0), \
543 | svg_attr("y", "%d", 0), \
544 | svg_attr("width", "%s", "100%"), \
545 | svg_attr("height", "%d", score), \
546 | svg_attr("stroke", "%s", "none"), \
547 | svg_attr("fill", "%s", color) \
548 | )
549 | score_back(max_score, "#ccebc5"); // green
550 | score_back(28, "#ffffcc"); // yellow
551 | score_back(20, "#fbb4ae"); // red
552 |
553 | /* Allocate 15 characters per point.
554 | 4 = max length is capped in kb range, will start compressing if larger
555 | 2 = '.5' added to point
556 | 1 = ','
557 | 5 = '##.##' for float average
558 | 1 = ' '
559 | 2 = padding for miscalculation
560 | */
561 | size_t mean_line_points_length = 15*data->max_length;
562 | char * mean_line_points = malloc(mean_line_points_length);
563 |
564 | /* Make line start off graph */
565 | snprintf(mean_line_points, mean_line_points_length, "0,%0.2f ", averages[0]);
566 |
567 | for (x = 0; x < data->max_length; x++) {
568 | for (y = 0; y < max_score; y++) {
569 | if(data->bases[x].scores[y+offset] > 0)
570 | svg_simple_tag("rect", 8,
571 | svg_attr("x", "%d", x),
572 | svg_attr("y", "%d", y),
573 | svg_attr("fill-opacity", "%f", (float)(data->bases[x].scores[y+offset])/100.0),
574 | svg_attr("width", "%d", 1),
575 | svg_attr("height", "%d", 1),
576 | svg_attr("stroke", "%s", "none"),
577 | svg_attr("stroke-width", "%d", 0),
578 | svg_attr("fill", "%s", "black")
579 | );
580 | }
581 |
582 | snprintf(tmp, 20, "%d.5,%0.2f ", x, averages[x]);
583 | strncat(mean_line_points, tmp, mean_line_points_length);
584 | }
585 |
586 | /* Make line end off graph */
587 | snprintf(tmp, 20, "%d,%0.2f", data->max_length, averages[data->max_length-1]);
588 | strncat(mean_line_points, tmp, mean_line_points_length);
589 |
590 |
591 | /* Print mean line */
592 | svg_simple_tag("polyline", 6,
593 | svg_attr("points", "%s", mean_line_points),
594 | svg_attr("stroke", "%s", "black"),
595 | svg_attr("stroke-width", "%f", 0.5),
596 | svg_attr("stroke-opacity", "%f", 0.5),
597 | svg_attr("fill", "%s", "none"),
598 | svg_attr("stroke-linejoin", "%s", "round")
599 | );
600 |
601 | free(mean_line_points);
602 |
603 | svg_end_tag("svg"); // Heatmap
604 | svg_end_tag("g"); // Heatmap
605 |
606 |
607 | /* Lables */
608 |
609 | svg_start_tag("text", 5,
610 | svg_attr("y", "%d", 350),
611 | svg_attr("fill", "%s", "#888"),
612 | svg_attr("x", "%d", 5),
613 | svg_attr("font-family", "%s", "sans-serif"),
614 | svg_attr("font-size", "%s", "15px")
615 | );
616 | printf("%s\n", "Per Base Sequence Quality");
617 | svg_end_tag("text");
618 |
619 |
620 | /* Print base quality labels in center, only needed the first time
621 | 465 = width of vertical section (450) + width of margin (30) halved
622 | 112 = start of graph (105) + half size of text (7)
623 | */
624 | if(position==0){
625 | svg_center_label(465, 112, "#888", "%d", max_score);
626 | svg_center_label(465, 112+(int)((max_score-28)*250/max_score), "#888", "%d", 28);
627 | svg_center_label(465, 112+(int)((max_score-20)*250/max_score), "#888", "%d", 20);
628 | }
629 |
630 |
631 | /*************** Length Distro ***************/
632 |
633 | /* Length Distro graph grows away from heatmap. No need to flip or have
634 | negative y*/
635 | svg_start_tag("svg", 6,
636 | svg_attr("x", "%d", 0),
637 | svg_attr("y", "%d", 360),
638 | svg_attr("width", "%d", 450),
639 | svg_attr("height", "%d", 100),
640 | svg_attr("preserveAspectRatio", "%s", "none"),
641 | svg_attr("viewBox", "0 0 %d 100", data->max_length)
642 | );
643 |
644 | /* Set background color */
645 | svg_simple_tag("rect", 3,
646 | svg_attr("width", "%s", "100%"),
647 | svg_attr("height", "%s", "100%"),
648 | svg_attr("fill", "%s", "#EEE")
649 | );
650 |
651 | for (x = 0; x < data->max_length; x++) {
652 | if( data->bases[x].length_count > 0)
653 | svg_simple_tag("rect", 6,
654 | svg_attr("x", "%d", x),
655 | svg_attr("y", "%d", 0),
656 | svg_attr("width", "%d", 1),
657 | svg_attr("height", "%d", data->bases[x].length_count),
658 | svg_attr("stroke", "%s", "none"),
659 | svg_attr("fill", "%s", "steelblue")
660 | );
661 | }
662 |
663 |
664 |
665 | svg_end_tag("svg"); // Length Distro
666 |
667 |
668 | /* Lables */
669 |
670 | svg_start_tag("text", 5,
671 | svg_attr("y", "%d", 455),
672 | svg_attr("fill", "%s", "#888"),
673 | svg_attr("x", "%d", 5),
674 | svg_attr("font-family", "%s", "sans-serif"),
675 | svg_attr("font-size", "%s", "15px")
676 | );
677 | printf("%s\n", "Length Distribution");
678 | svg_end_tag("text");
679 |
680 | if(position == 0){
681 | svg_axis_label(-410, -5, -90, "Percent");
682 | svg_axis_number(-5, 370, "end", 0);
683 | svg_axis_number(-5, 460, "end", 100);
684 | }else{
685 | svg_axis_label(410, -455, 90, "Percent");
686 | svg_axis_number(455, 370, "start", 0);
687 | svg_axis_number(455, 460, "start", 100);
688 | }
689 |
690 |
691 | /*************** Adapter Distro ***************/
692 |
693 | if(adapters_used==1){
694 | /* Adapter Distro graph grows away from heatmap. No need to flip or have
695 | negative y*/
696 | svg_start_tag("svg", 6,
697 | svg_attr("x", "%d", 0),
698 | svg_attr("y", "%d", 465),
699 | svg_attr("width", "%d", 450),
700 | svg_attr("height", "%d", 100),
701 | svg_attr("preserveAspectRatio", "%s", "none"),
702 | svg_attr("viewBox", "0 0 %d 100", data->max_length)
703 | );
704 |
705 | /* Set background color */
706 | svg_simple_tag("rect", 3,
707 | svg_attr("width", "%s", "100%"),
708 | svg_attr("height", "%s", "100%"),
709 | svg_attr("fill", "%s", "#EEE")
710 | );
711 |
712 | for (x = 0; x < data->max_length; x++) {
713 | if( data->bases[x].kmer_count > 0)
714 | svg_simple_tag("rect", 6,
715 | svg_attr("x", "%d", x),
716 | svg_attr("y", "%d", 0),
717 | svg_attr("width", "%d", 1),
718 | svg_attr("height", "%d", data->bases[x].kmer_count),
719 | svg_attr("stroke", "%s", "none"),
720 | svg_attr("fill", "%s", "steelblue")
721 | );
722 | }
723 |
724 | svg_end_tag("svg"); // Adapter Distro
725 |
726 | /* Lables */
727 |
728 | svg_start_tag("text", 5,
729 | svg_attr("y", "%d", 560),
730 | svg_attr("fill", "%s", "#888"),
731 | svg_attr("x", "%d", 5),
732 | svg_attr("font-family", "%s", "sans-serif"),
733 | svg_attr("font-size", "%s", "15px")
734 | );
735 | printf("%s\n", "Adapter Distribution");
736 | svg_end_tag("text");
737 |
738 | if(position == 0){
739 | svg_axis_label(-515, -5, -90, "Percent");
740 | svg_axis_number(-5, 475, "end", 0);
741 | svg_axis_number(-5, 565, "end", 100);
742 | }else{
743 | svg_axis_label(515, -455, 90, "Percent");
744 | svg_axis_number(455, 475, "start", 0);
745 | svg_axis_number(455, 565, "start", 100);
746 | }
747 | }
748 |
749 | /*************** Bottom Label ***************/
750 | y = 470;
751 | if(adapters_used==1) y+=105;
752 |
753 | svg_axis_label(225, y+5, 0, "Base Pairs");
754 | svg_axis_number(0, y, "middle", 0);
755 | svg_axis_number(450, y, "middle", data->max_length);
756 |
757 |
758 | svg_end_tag("g"); // rug plot vertical section
759 |
760 | /*************** Score Distro ***************/
761 |
762 | /* Score Distro graph is shown on either side of the heatmap, so it needs to
763 | flip vertically if position == 0. Flipped horizontally as well. Use
764 | negative positions in directions svg is flipped*/
765 |
766 | svg_start_tag("g", 1,
767 | svg_attr("transform", "translate(%d,%d) scale(%d, %d)",
768 | (position == 0)?125:1065, 355, (position == 0)?-1:1,-1));
769 |
770 | svg_start_tag("svg", 4,
771 | svg_attr("width", "%d", 100),
772 | svg_attr("height", "%d", 250),
773 | svg_attr("preserveAspectRatio", "%s", "none"),
774 | svg_attr("viewBox", "0 0 100 %d", max_score)
775 | );
776 |
777 | /* Set background color */
778 | svg_simple_tag("rect", 3,
779 | svg_attr("width", "%s", "100%"),
780 | svg_attr("height", "%s", "100%"),
781 | svg_attr("fill", "%s", "#EEE")
782 | );
783 |
784 | for (y = 0; y < max_score; y++) {
785 | if(total_counts[y] > 0)
786 | svg_simple_tag("rect", 6,
787 | svg_attr("x", "%d", 0),
788 | svg_attr("y", "%d", y),
789 | svg_attr("width", "%d", total_counts[y]/number_of_bases),
790 | svg_attr("height", "%d", 1),
791 | svg_attr("stroke", "%s", "none"),
792 | svg_attr("fill", "%s", "steelblue")
793 | );
794 | }
795 |
796 | svg_end_tag("svg"); // Score Distro
797 | svg_end_tag("g"); // Score Distro
798 |
799 | /* Lables */
800 |
801 | if(position == 0){
802 | svg_start_tag("text", 5,
803 | svg_attr("y", "%d", 335),
804 | svg_attr("fill", "%s", "#888"),
805 | svg_attr("x", "%d", 30),
806 | svg_attr("font-family", "%s", "sans-serif"),
807 | svg_attr("font-size", "%s", "15px")
808 | );
809 | svg_start_tag("tspan", 0);
810 | printf("%s\n", "Score");
811 | svg_end_tag("tspan");
812 |
813 | svg_start_tag("tspan", 2, svg_attr("dy", "%d", 15), svg_attr("x", "%d", 30));
814 | printf("%s\n", "Distribution");
815 | svg_end_tag("tspan");
816 |
817 | svg_end_tag("text");
818 |
819 | svg_axis_label(72, 100, 0, "Percent");
820 | /* svg_axis_number(125, 100, "middle", 0); */
821 | svg_axis_number(25, 100, "middle", 100);
822 |
823 | svg_axis_label(-230, 20, -90, "Score");
824 | svg_axis_number(20, 110, "end", max_score);
825 | svg_axis_number(20, 355, "end", 1);
826 |
827 | }else{
828 | svg_start_tag("text", 5,
829 | svg_attr("y", "%d", 335),
830 | svg_attr("fill", "%s", "#888"),
831 | svg_attr("x", "%d", 1070),
832 | svg_attr("font-family", "%s", "sans-serif"),
833 | svg_attr("font-size", "%s", "15px")
834 | );
835 | svg_start_tag("tspan", 0);
836 | printf("%s\n", "Score");
837 | svg_end_tag("tspan");
838 |
839 | svg_start_tag("tspan", 2, svg_attr("dy", "%d", 15), svg_attr("x", "%d", 1070));
840 | printf("%s\n", "Distribution");
841 | svg_end_tag("tspan");
842 |
843 | svg_end_tag("text");
844 |
845 | svg_axis_label(1115, 100, 0, "Percent");
846 | /* svg_axis_number(1070, 100, "middle", 0); */
847 | svg_axis_number(1165, 100, "middle", 100);
848 |
849 | svg_axis_label(230, -1170, 90, "Score");
850 | svg_axis_number(1170, 110, "start", max_score);
851 | svg_axis_number(1170, 355, "start", 1);
852 | }
853 |
854 | svg_end_tag("g"); // rug plot
855 |
856 | }
857 |
858 | int main (int argc, char **argv)
859 | {
860 | struct arguments arguments;
861 | arguments = parse_options(argc, argv);
862 |
863 | int paired, unpaired, adapters;
864 | int width, height;
865 | int *kmers = NULL;
866 |
867 | paired = (arguments.forward != NULL && arguments.reverse != NULL);
868 | unpaired = (arguments.unpaired != NULL);
869 | adapters = (arguments.adapters != NULL);
870 |
871 | /* If paired and unparied data are both set or unset, then throw error */
872 | if(paired == unpaired){
873 | printf("%s\n", "Usage: quack [OPTION...]\nTry `quack --help' or `quack --usage' for more information.");
874 | exit(1);
875 | }
876 |
877 | if(adapters) kmers = read_adapters(arguments.adapters);
878 |
879 | width = (paired)?1195:615;
880 | height = (adapters)?610:510;
881 |
882 | if(arguments.name != NULL)
883 | height += 30;
884 |
885 | svg_start_tag("svg", 5,
886 | svg_attr("width", "%d", width),
887 | svg_attr("height", "%d", height),
888 | svg_attr("viewBox", "%d %d %d %d", 0, 0, width, height),
889 | svg_attr("xmlns", "%s", "http://www.w3.org/2000/svg"),
890 | svg_attr("xmlns:xlink", "%s", "http://www.w3.org/1999/xlink")
891 | );
892 |
893 | // If name is given, add to middle of viewBox (half of width + min-x of viewbox)
894 | if(arguments.name != NULL){
895 | svg_start_tag("text", 6,
896 | svg_attr("x", "%d", (width/2)),
897 | svg_attr("y", "%d", 30),
898 | svg_attr("font-family", "%s", "sans-serif"),
899 | svg_attr("text-anchor", "%s", "middle"),
900 | svg_attr("font-size", "%s", "30px"),
901 | svg_attr("fill", "%s", "black"));
902 | printf("%s", arguments.name);
903 | svg_end_tag("text");
904 |
905 | svg_start_tag("g", 1,
906 | svg_attr("transform", "translate(%d %d)", 0, 30)
907 | );
908 |
909 | }
910 |
911 | sequence_data *data = read_fastq(((paired)?arguments.forward:arguments.unpaired), kmers);
912 | sequence_data *transformed_data = transform(data);
913 | draw(transformed_data, 0, adapters);
914 | free(data);
915 |
916 | if(paired){
917 | data = read_fastq(arguments.reverse, kmers);
918 | transformed_data = transform(data);
919 | draw(transformed_data, 1, adapters);
920 | free(data);
921 | }
922 |
923 | if(arguments.name != NULL) svg_end_tag("g");
924 |
925 | svg_end_tag("svg");
926 |
927 | exit (0);
928 | }
929 |
--------------------------------------------------------------------------------
/svg.c:
--------------------------------------------------------------------------------
1 | #include "svg.h"
2 |
3 | #include
4 | #include
5 | #include
6 | #include
7 |
8 | #define INDENT " "
9 | int _svg_indent_level = 0;
10 |
11 |
12 | char* svg_attr(const char* name, const char* fmt, ...){
13 | char *attr;
14 | int retval, length, cur_length;
15 | va_list vl;
16 |
17 | /* Use vsnprintf to get the length of value string before allocating memory*/
18 | va_start(vl, fmt);
19 | retval = vsnprintf(NULL, 0, fmt, vl);
20 | va_end(vl);
21 | if(retval < 0) return NULL;
22 |
23 | /* Allocate enough memory for entire attribute string:
24 | ' $name="$value"'
25 | length($name) + length($value) + 4 literals [ =""] + null char
26 | */
27 | length = retval + strlen(name) + 5;
28 | attr = malloc(length);
29 | if(attr == NULL) return NULL;
30 |
31 |
32 | /* Print beginning of attribute: ' $name="' ; freeing memory if failed*/
33 | retval = snprintf(attr, length-1, " %s=\"", name);
34 | if(retval < 0){
35 | free(attr);
36 | return NULL;
37 | }
38 | cur_length = retval;
39 |
40 | /* Print attribute value; freeing memory if failed*/
41 | va_start(vl, fmt);
42 | retval = vsnprintf(attr + cur_length, length-cur_length-1, fmt, vl);
43 | va_end(vl);
44 | if(retval < 0){
45 | free(attr);
46 | return NULL;
47 | }
48 | cur_length += retval;
49 |
50 | /* Close quote and terminate string */
51 | attr[cur_length] = '"';
52 | attr[cur_length+1] = '\0';
53 |
54 | return attr;
55 | }
56 |
57 |
58 |
59 | void _svg_tag(const int simple, const char* type, int num, ...){
60 | int i = 0;
61 | va_list vl;
62 | char* attr;
63 |
64 | /* Indent current tag */
65 | for( i = 0; i < _svg_indent_level; i++)
66 | printf(INDENT);
67 |
68 | /* Increase indent level if started tag with internal elements */
69 | if(!simple)
70 | _svg_indent_level++;
71 |
72 | /* Open tag */
73 | printf("<%s", type);
74 |
75 | /* Print each attribute, free memory as used */
76 | va_start(vl, num);
77 | for(i = 0; i < num; i++){
78 | attr = va_arg(vl, char*);
79 | printf("%s", attr);
80 | free(attr);
81 | }
82 | va_end(vl);
83 |
84 | /* print nested closing tag if no internal elements expected */
85 | if(simple) printf("/");
86 |
87 | /* close tag */
88 | printf(">\n");
89 | }
90 |
91 |
92 | void svg_end_tag(const char* type){
93 | int i = 0;
94 |
95 | if(_svg_indent_level > 0)
96 | _svg_indent_level--;
97 |
98 | for( i = 0; i < _svg_indent_level; i++)
99 | printf(INDENT);
100 |
101 |
102 | printf("%s>\n", type);
103 |
104 | }
105 |
--------------------------------------------------------------------------------
/svg.h:
--------------------------------------------------------------------------------
1 | #ifndef __SVG_H
2 | #define __SVG_H
3 |
4 | /* Add an attribute to the svg tag.
5 | - `name` = name of attribute.
6 | - `fmt` = printf format
7 | - `...` = are variables passed to printf
8 | */
9 | /* #define svg_attr(name, type, ...) " " #name "=" #type , __VA_ARGS__ */
10 | char* svg_attr(const char* name, const char* fmt, ...);
11 |
12 | /* Print an svg tag (i.e ). `svg_start_tag` doesn't add a final '/',
13 | while `svg_simple_tag` does.
14 | - `type` = type of element (e.g. 'svg', 'line', 'rect', etc)
15 | - `num` = number of attributes
16 | - `...` = a list of svg_attr
17 | */
18 | void _svg_tag(const int simple, const char* type, int num, ...);
19 | #define svg_start_tag(...) _svg_tag(0, __VA_ARGS__)
20 | #define svg_simple_tag(...) _svg_tag(1, __VA_ARGS__)
21 |
22 |
23 | /* Prints closing svg tag (i.e ).
24 | - `type` = type of element (e.g. 'svg', 'line', 'rect', etc)
25 | */
26 | void svg_end_tag(const char* type);
27 |
28 |
29 | #endif
30 |
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