archive:bioinformatic_tools:quake
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archive:bioinformatic_tools:quake [2011/05/07 11:53] eyliaw |
archive:bioinformatic_tools:quake [2015/07/28 06:26] (current) ceisenhart ↷ Page moved from bioinformatic_tools:quake to archive:bioinformatic_tools:quake |
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| Finally, correct the reads: | Finally, correct the reads: | ||
| - | correct -f [fastq list file] -k [k-mer size] -c [cutoff] -m [counts file] -p [number of cores] | + | correct -f [fastq file list] -k [k-mer size] -c [cutoff] -m [counts file] -p [number of cores] -z (gzips the output) |
| - | [Kevin] I think that we want to select the k-mer size manually, rather than relying on quake. Their default cutoff is very conservative, and we'll probably do better over-correcting than under-correcting. | + | In the file list, you should tab-separate paired end reads. Also, be sure that all .'s in the sequence are written as N's. If the quality scores are written as Phred + 64, you can use -q 64 to handle it. |
| - | If we look at the hugely over-represented k-mers (like the adapter sequences), and compare the true sequences to one that are one base different, we see that the true ones are about 30 times as frequent. Thus quake's idea of correcting only the rarely seen k-mers isn't quite right. What we really want to correct are those k-mers that are close neighbors of much more frequent k-mers. I've not figured out yet precisely what "much more frequent" should mean. | + | |
| + | K-mer counts can also be pre-filtered to save space. | ||
| - | ---- | + | Quake dev: |
| - | Can the k-mer counts be pre-filtered to save space? | + | Once you've decided on a cutoff, Quake ignores all of the k-mers below that cutoff. So sure, you can filter the file to save some disk space. But having all of the k-mer counts is best for choosing the cutoff. My cov_model.py script to automatically choose the cutoff requires them. |
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| + | [Kevin] I think that we want to select the k-mer size manually, rather than relying on quake. Their default cutoff is very conservative, and we'll probably do better over-correcting than under-correcting. | ||
| + | If we look at the hugely over-represented k-mers (like the adapter sequences), and compare the true sequences to one that are one base different, we see that the true ones are about 30 times as frequent. Thus quake's idea of correcting only the rarely seen k-mers isn't quite right. What we really want to correct are those k-mers that are close neighbors of much more frequent k-mers. I've not figured out yet precisely what "much more frequent" should mean. | ||
| - | Well sort of. Once you've decided on a cutoff, Quake ignores all of the k-mers below that cutoff. So sure, you can filter the file to save some disk space. But having all of the k-mer counts is best for choosing the cutoff. My cov_model.py script to automatically choose the cutoff requires them. | + | ===== Potential Problems ====== |
| - | ---- | + | * Input files need to have an extension, or Quake will throw a substr error when trying to merge hidden files into a result. |
| + | * With paired-end input, Quake will output two files for each paired-end read. One will be the cor.fastq file, which contains corrected, paired reads. The other will be the cor_single.fastq file, which contains reads where only one pair could be corrected. You can treat the cor_single.fastq file as a single read file. | ||
| ===== Methods ===== | ===== Methods ===== | ||
archive/bioinformatic_tools/quake.1304769198.txt.gz · Last modified: 2011/05/07 11:53 by eyliaw
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