Question: Re: Cufflinks With Reference Annotation And Without Reference Annotation
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gravatar for Crystal Goh
7.3 years ago by
Crystal Goh • 10
Crystal Goh • 10 wrote:
Dear Jeremy, Thanks. I am sorry for sending mutiple emails. I will email only once next time. Thank you. Best regards, Crystal CC: galaxy-user@lists.bx.psu.edu; jen@bx.psu.edu; dbschak@nus.edu.sg To: crystal86@live.com.my Subject: Re: Cufflinks with reference annotation and without reference annotation Date: Wed, 17 Aug 2011 14:17:26 -0400 Crystal, If you provide a gene annotation to Cufflinks, the transcripts produced will match those in the annotation exactly. If you assemble without a gene annotation, the transcripts produced will match the reference in some cases, but, in others, will not match the reference due to small and/or large errors. Because '=' denotes an exact match between an assembled transcript and a reference transcript, more '=' are to be expected when Cufflinks has a gene annotation. Finally, a couple procedural issues: *please send questions about analyses and tool usage to the galaxy- user mailing list, not galaxy-dev or individual developers; *please do not send duplicate emails as it can confuse our tracking system and slow down our response rather than speed it up. Good luck, J. Hi, I am Crystal. I have some problem with Cuffdiff output. Hope can get some advice. Thanks. After aligning RNA-seq reads with Tophat, I used the Tophat output for Cufflinks. For Cufflinks, I tried two approaches and compared the results: 1st approach: Put zebrafish Ensembl GTF as reference annotation 2nd approach: without reference annotation. Attached word document is the workflow and parameters I set for these 2 approaches. When I compared the output of Cuffdiff between these 2 approaches, a total of 48584 tracking id with class code "=" was observed in trancript FPKM tracking file from Approach 1, whereas there is only 1248 tracking id with class code '=' from Approach 2 (I attached transcript FPKM tracking files from approach 1 and 2) In my opinion, I should observe 48584 tracking id with class code '=' and additional tracking id with other class codes in transcript FPKM tracking file from Approach 2. Can I get advice on this? Thank you. Best regards, Crystal <workflow and="" parameter="" for="" 2="" approaches.zip=""><approach 1="" transcript="" fpkm="" tracking="" (cufflinks="" with="" reference="" annotation).zip=""><approach 2="" transcript="" fpkm="" tracking="" (cufflinks="" without="" reference="" annotation).zip="">
rna-seq cufflinks • 2.9k views
ADD COMMENT • link • modified 7.3 years ago by Jeremy Goecks • 2.2k • written 7.3 years ago by Crystal Goh • 10
0
gravatar for Jeremy Goecks
7.3 years ago by
Jeremy Goecks • 2.2k
Jeremy Goecks • 2.2k wrote:
Crystal, If you provide a gene annotation to Cufflinks, the transcripts produced will match those in the annotation exactly. If you assemble without a gene annotation, the transcripts produced will match the reference in some cases, but, in others, will not match the reference due to small and/or large errors. Because '=' denotes an exact match between an assembled transcript and a reference transcript, more '=' are to be expected when Cufflinks has a gene annotation. Finally, a couple procedural issues: *please send questions about analyses and tool usage to the galaxy- user mailing list, not galaxy-dev or individual developers; *please do not send duplicate emails as it can confuse our tracking system and slow down our response rather than speed it up. Good luck, J.
ADD COMMENT • link written 7.3 years ago by Jeremy Goecks • 2.2k
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