1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
148
149
150
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
226
227
228
229
230
231
232
233
234
235
236
237
238
239
240
241
242
243
244
245
246
247
248
249
250
251
252
253
254
255
256
257
258
259
260
261
262
263
264
265
266
267
268
269
270
271
272
273
274
275
276
277
278
279
280
281
282
283
284
285
286
287
288
289
290
291
292
293
294
295
296
297
298
299
300
301
302
303
304
305
306
307
308
309
extern crate fasten;
extern crate fastq;
extern crate getopts;
extern crate rand;
use std::io::stdin;
use std::collections::HashMap;
use std::env;
use std::f32;
use fasten::fasten_base_options;
use fastq::{Parser, Record};
use fasten::logmsg;
const TEN: f32 = 10.0;
const READ_SEPARATOR :char = '~';
fn main(){
let args: Vec<String> = env::args().collect();
let mut opts = fasten_base_options();
let default_phred_min_char:char = '!';
let default_phred_max_char:char = 'I';
let default_phred_min:u8 = default_phred_min_char as u8 - 33;
let default_phred_max:u8 = default_phred_max_char as u8 - 33;
let mut qual_range_string = String::with_capacity((default_phred_max - default_phred_min + 1) as usize);
for phred in default_phred_min..default_phred_max+1 {
qual_range_string.push((phred+33) as char);
}
opts.optopt("","max-qual-char",
format!("Maximum quality character (default: {})", default_phred_max_char).as_str(),
"CHAR");
opts.optopt("","min-qual-char",
format!("Minimum quality character (default: {})", default_phred_min_char).as_str(),
"CHAR");
let matches = opts.parse(&args[1..]).expect("Parsing parameters");
if matches.opt_present("h") {
println!("Collapse identical reads into single reads, recalculating quality values. If paired end, then each set of reads must be identical to be collapsed. Warning: due to multiple reads collapsing into one, read identifiers will be reconstituted.");
println!("{}",opts.usage(&opts.short_usage(&args[0])));
println!("NOTE: range of quality scores is {}", qual_range_string);
std::process::exit(0);
}
let max_qual_char:char = matches.opt_default("max-qual-char", &default_phred_max_char.to_string())
.unwrap_or(String::from(default_phred_max_char))
.parse()
.expect("ERROR converting --max-qual-int value to integer");
let mut min_qual_char:char =
matches.opt_default("min-qual-char", &default_phred_min_char.to_string())
.unwrap_or(String::from(default_phred_min_char))
.parse()
.expect("ERROR converting --min-qual-int value to integer");
if min_qual_char < default_phred_min_char {
logmsg("--min-qual-char was less than the default minimum and so it will be set to the default");
min_qual_char = default_phred_min_char;
}
let min_qual:u8 = min_qual_char as u8;
let max_qual:u8 = max_qual_char as u8;
let paired_end = matches.opt_present("paired-end");
let _num_cpus:usize = {
if matches.opt_present("numcpus") {
logmsg("Warning: This script does not make use of --numcpus");
1 as usize
} else {
1 as usize
}
};
let mut seq_count :HashMap<String, u32> =HashMap::new();
let mut seq_error_rate :HashMap<String, Vec<f32>> = HashMap::new();
let parser = Parser::new(stdin());
let mut parser_getter = parser.ref_iter();
parser_getter.advance().expect("Could not read the first fastq entry");
while let Some(record1) = parser_getter.get() {
let mut id:Vec<u8> = record1.head().to_vec();
let mut seq:Vec<u8> = record1.seq().to_vec();
let mut qual:Vec<u8> = record1.qual().to_vec();
if paired_end {
match parser_getter.advance() {
Ok(_) => {},
Err(err) => {
panic!("ERROR: could not read the second entry in a paired end read: {}", err);
}
};
let record2 = &parser_getter.get().expect("ERROR parsing second pair in a paired end read");
let id2:&[u8] = record2.head();
let seq2:&[u8] = record2.seq();
let qual2:&[u8]= record2.qual();
id.push(READ_SEPARATOR as u8);
seq.push(READ_SEPARATOR as u8);
qual.push(READ_SEPARATOR as u8);
id.extend_from_slice(id2);
seq.extend_from_slice(seq2);
qual.extend_from_slice(qual2);
}
let seq_string:String = String::from(
std::str::from_utf8(&seq[..])
.expect("ERROR converting slice to str")
);
let count = seq_count.entry(seq_string.clone()).or_insert(0);
*count += 1 as u32;
if !seq_error_rate.contains_key(&seq_string) {
let mut qual_vec:Vec<f32> = vec![];
for q in qual {
if q == READ_SEPARATOR as u8 {
qual_vec.push(q as u8 as f32);
continue;
}
let qual_int = q as u8 as f32 - 33.0;
let p :f32 = TEN.powf((-1.0 * qual_int)/TEN);
qual_vec.push(p);
}
seq_error_rate.insert(seq_string.clone(), qual_vec);
}
else {
let qual_vec = seq_error_rate.entry(seq_string.clone()).or_insert(Vec::new());
let these_errors = qual.into_iter().map(|qual_char|{
if qual_char == READ_SEPARATOR as u8 {
return qual_char as u8 as f32;
}
let qual_int = qual_char as u8 as f32 - 33.0;
let p :f32 = TEN.powf((-1.0 * qual_int)/TEN);
return p;
}).collect();
let new_qual = combine_error_vectors(&qual_vec,&these_errors);
*qual_vec = new_qual;
}
match &parser_getter.advance() {
Ok(_) => {},
Err(_) => {break;}
};
}
let mut seq_counter=0;
for (seq,combined_qual) in seq_error_rate {
seq_counter += 1;
let mut qual_cigar = String::new();
for p in combined_qual {
let mut qual_recalc :f32 = -TEN * (p).log(TEN)+33.0;
if qual_recalc.is_infinite() || qual_recalc > max_qual as f32 {
qual_recalc = max_qual as f32;
}
let mut qual_recalc_char = qual_recalc.round() as u8 as char;
if (qual_recalc_char as u8) > max_qual {
qual_recalc_char = max_qual_char;
}
if (qual_recalc_char as u8) < min_qual {
qual_recalc_char = min_qual_char;
}
qual_cigar.push(qual_recalc_char);
}
if paired_end {
let separator_pos = seq.find(READ_SEPARATOR).expect("ERROR finding read separator");
let r1_seq = seq[0..separator_pos].to_string();
let r2_seq = seq[separator_pos+1..].to_string();
let r1_qual= qual_cigar[0..separator_pos].to_string();
let r2_qual= qual_cigar[separator_pos+1..].to_string();
println!("@{}/1\n{}\n+\n{}",seq_counter,r1_seq,r1_qual);
println!("@{}/2\n{}\n+\n{}",seq_counter,r2_seq,r2_qual);
} else {
println!("@{}\n{}\n+\n{}",seq_counter,seq,qual_cigar);
}
}
}
fn combine_error_vectors(errors1 :&Vec<f32>, errors2: &Vec<f32>) -> Vec<f32> {
if errors1.len() != errors2.len() {
panic!("Lengths of error vectors do not match: {} and {}", errors1.len(), errors2.len());
}
let mut errors_iter2=errors2.iter();
let mut new_errors :Vec<f32> = Vec::new();
for p1 in errors1 {
let p2 = errors_iter2.next().expect("ERROR: could not get the error probability from the second read");
new_errors.push(p1 * p2);
}
return new_errors;
}