Agent skill
bio-fastq-quality
Work with FASTQ quality scores using Biopython. Use when analyzing read quality, filtering by quality, trimming low-quality bases, or generating quality reports.
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npx add-skill https://github.com/majiayu000/claude-skill-registry/tree/main/skills/data/fastq-quality
SKILL.md
FASTQ Quality Scores
Analyze and manipulate FASTQ quality scores using Biopython.
Required Imports
python
from Bio import SeqIO
from Bio.Seq import Seq
Accessing Quality Scores
Quality scores are stored in letter_annotations['phred_quality'] as a list of integers.
python
for record in SeqIO.parse('reads.fastq', 'fastq'):
qualities = record.letter_annotations['phred_quality']
print(record.id, qualities[:10]) # First 10 quality scores
Quality Score Basics
| Phred Score | Error Probability | Accuracy |
|---|---|---|
| 10 | 1 in 10 | 90% |
| 20 | 1 in 100 | 99% |
| 30 | 1 in 1000 | 99.9% |
| 40 | 1 in 10000 | 99.99% |
Code Patterns
Calculate Average Quality per Read
python
for record in SeqIO.parse('reads.fastq', 'fastq'):
quals = record.letter_annotations['phred_quality']
avg_qual = sum(quals) / len(quals)
print(f'{record.id}: {avg_qual:.1f}')
Filter Reads by Mean Quality
python
def high_quality_reads(records, min_avg_qual=20):
for record in records:
quals = record.letter_annotations['phred_quality']
if sum(quals) / len(quals) >= min_avg_qual:
yield record
records = SeqIO.parse('reads.fastq', 'fastq')
good_reads = high_quality_reads(records, min_avg_qual=25)
SeqIO.write(good_reads, 'filtered.fastq', 'fastq')
Filter by Minimum Quality at Any Position
python
def all_bases_above(records, min_qual=20):
for record in records:
if min(record.letter_annotations['phred_quality']) >= min_qual:
yield record
Trim Low-Quality Ends (3' Trimming)
python
def trim_low_quality(record, min_qual=20):
quals = record.letter_annotations['phred_quality']
trim_pos = len(quals)
for i in range(len(quals) - 1, -1, -1):
if quals[i] >= min_qual:
trim_pos = i + 1
break
return record[:trim_pos]
records = SeqIO.parse('reads.fastq', 'fastq')
trimmed = (trim_low_quality(rec) for rec in records)
SeqIO.write(trimmed, 'trimmed.fastq', 'fastq')
Sliding Window Quality Trim
python
def sliding_window_trim(record, window_size=5, min_avg_qual=20):
quals = record.letter_annotations['phred_quality']
for i in range(len(quals) - window_size + 1):
window = quals[i:i + window_size]
if sum(window) / window_size < min_avg_qual:
return record[:i] if i > 0 else None
return record
Quality Statistics Summary
python
import statistics
all_quals = []
for record in SeqIO.parse('reads.fastq', 'fastq'):
all_quals.extend(record.letter_annotations['phred_quality'])
print(f'Mean quality: {statistics.mean(all_quals):.1f}')
print(f'Median quality: {statistics.median(all_quals):.1f}')
print(f'Min quality: {min(all_quals)}')
print(f'Max quality: {max(all_quals)}')
Per-Position Quality Profile
python
from collections import defaultdict
position_quals = defaultdict(list)
for record in SeqIO.parse('reads.fastq', 'fastq'):
for i, q in enumerate(record.letter_annotations['phred_quality']):
position_quals[i].append(q)
for pos in sorted(position_quals.keys())[:20]:
quals = position_quals[pos]
print(f'Position {pos}: mean={sum(quals)/len(quals):.1f}')
Count Reads by Quality Threshold
python
thresholds = [20, 25, 30, 35]
counts = {t: 0 for t in thresholds}
for record in SeqIO.parse('reads.fastq', 'fastq'):
avg = sum(record.letter_annotations['phred_quality']) / len(record.seq)
for t in thresholds:
if avg >= t:
counts[t] += 1
for t, c in counts.items():
print(f'Q>={t}: {c} reads')
Remove N Bases and Low Quality Together
python
def clean_read(record, min_qual=20):
seq = str(record.seq)
quals = record.letter_annotations['phred_quality']
keep = [(s, q) for s, q in zip(seq, quals) if s != 'N' and q >= min_qual]
if not keep:
return None
new_seq, new_quals = zip(*keep)
new_record = record[:0] # Empty copy with same metadata
new_record.seq = Seq(''.join(new_seq))
new_record.letter_annotations['phred_quality'] = list(new_quals)
return new_record
FASTQ Format Variants
| Variant | Format String | Quality Encoding | ASCII Range |
|---|---|---|---|
| Sanger/Illumina 1.8+ | 'fastq' |
Phred+33 (standard) | 33-126 |
| Solexa | 'fastq-solexa' |
Solexa+64 | 59-126 |
| Illumina 1.3-1.7 | 'fastq-illumina' |
Phred+64 | 64-126 |
Most modern data uses standard 'fastq' (Sanger/Illumina 1.8+).
Quality Score Conversion
For legacy data using different quality encodings:
python
from Bio.SeqIO.QualityIO import phred_quality_from_solexa, solexa_quality_from_phred
Convert Solexa to Phred
python
from Bio.SeqIO.QualityIO import phred_quality_from_solexa
# Convert single score
solexa_score = 10
phred_score = phred_quality_from_solexa(solexa_score)
# Convert list of scores
solexa_scores = [10, 20, 30, 40]
phred_scores = [phred_quality_from_solexa(s) for s in solexa_scores]
Convert Phred to Solexa
python
from Bio.SeqIO.QualityIO import solexa_quality_from_phred
phred_score = 30
solexa_score = solexa_quality_from_phred(phred_score)
Convert Between FASTQ Variants
python
from Bio import SeqIO
# Read old Illumina format, write standard format
records = SeqIO.parse('old_reads.fastq', 'fastq-illumina')
SeqIO.write(records, 'standard_reads.fastq', 'fastq')
# Read Solexa format, write standard format
records = SeqIO.parse('solexa_reads.fastq', 'fastq-solexa')
SeqIO.write(records, 'standard_reads.fastq', 'fastq')
Auto-Detect Quality Encoding
python
def detect_quality_encoding(filepath, sample_size=1000):
'''Guess FASTQ quality encoding from ASCII values'''
min_qual = 126
max_qual = 0
count = 0
with open(filepath) as f:
for i, line in enumerate(f):
if i % 4 == 3: # Quality line
for char in line.strip():
min_qual = min(min_qual, ord(char))
max_qual = max(max_qual, ord(char))
count += 1
if count >= sample_size:
break
if min_qual < 59:
return 'fastq' # Sanger/Illumina 1.8+ (Phred+33)
elif min_qual < 64:
return 'fastq-solexa' # Solexa+64
else:
return 'fastq-illumina' # Illumina 1.3+ (Phred+64)
Common Errors
| Error | Cause | Solution |
|---|---|---|
KeyError: 'phred_quality' |
Not FASTQ or wrong variant | Check format, try 'fastq-illumina' |
| Quality scores all 0 | Wrong encoding assumed | Try different FASTQ variant |
| Trimmed reads empty | Too aggressive trimming | Lower quality threshold |
Related Skills
- read-sequences - Parse FASTQ files
- filter-sequences - Filter reads by other criteria (length, content)
- paired-end-fastq - Handle R1/R2 paired quality filtering
- sequence-statistics - Generate summary statistics including quality
- alignment-files - After filtering, align reads with bwa/bowtie2; quality scores in BAM
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