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archive:bioinformatic_tools:quake [2011/04/24 20:55]
eyliaw [Running Quake]
archive:bioinformatic_tools:quake [2011/05/27 21:41]
eyliaw
Line 15: Line 15:
 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) 
 + 
 +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. 
 + 
 +K-mer counts can also be pre-filtered to save space. 
 + 
 +Quake dev:   
 +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. 
 + 
 + 
 +[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. 
 + 
 +===== 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.txt · Last modified: 2015/07/28 06:26 by ceisenhart