Transcriptome Coverage- Method Of Calculation
2
2
Entering edit mode
11.6 years ago
nayaganviji ▴ 20

how we calculated transcriptome coverage of non model species like Crow. i am plan to use GS FLX+ system for RNA sequencing.

rna seq • 5.9k views
ADD COMMENT
2
Entering edit mode
11.6 years ago
JC 13k

It's really complex, because transcripts are not distributed proportionally. Many genes will be expressed in 10-100's copies but very few will be in the order of 1000's and others in <10's, so a minimal coverage should include low expressed genes but you will have to reduce the highly expressed genes to maximize your chance to get them in the sequencing. Besides, the non-model specie makes this harder to estimate. Additionally, the sample is also a factor, in humans, the brain express a more diversity range of transcripts than other tissues.

*I almost mark this question as duplicate of your last one, but the question is more specific. Maybe you can try to combine your questions because they are related.

ADD COMMENT
1
Entering edit mode
11.6 years ago
Ketil 4.1k

To calculate average coverage, you take the total amount of sequence, and divide by the total size of the transcriptome. If you don't know the latter, you can get an estimate by clustering the reads, but this will have huge opportunities for error.

Minimum coverage is likely zero (many transcripts will not be expressed), and maximum coverage is typically some ribosomal RNAs, which can be 30-50% of your reads.

ADD COMMENT

Login before adding your answer.

Traffic: 2749 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6