SEQanswers

SEQanswers (http://seqanswers.com/forums/index.php)
-   Oxford Nanopore (http://seqanswers.com/forums/forumdisplay.php?f=45)
-   -   Using the rapid sequencing kit for cDNA (http://seqanswers.com/forums/showthread.php?t=102111)

jgibbons1 11-19-2021 10:51 AM

Using the rapid sequencing kit for cDNA
 
We haven't done RNA-seq with the minion yet, but have an extra Rapid Sequencing kit lying around. I am wondering if there's anything "wrong" with creating our own cDNA and then sequencing it with the Rapid Kit, rather than using the one of the cDNA Sequencing Kits.

We are interested in some isoforms between two samples and thought this might be a creative solution with the kit we have in hand.

Any suggestions would be appreciated!

Thanks.

jgibbons1 11-29-2021 05:12 AM

To answer my own question, after thinking about this more...

This kit includes a fragmentation step, which would result in cDNA fragmentation and limit sequencing cDNA of full in tact isoforms. There's a risk the isoforms we're interested in will be fragmented.

gringer 01-18-2022 06:23 PM

Also, the cDNA kits are a stranded sequencing process (different adapter sequences for forward and reverse); you'd lose that benefit by using either the ligation kit or rapid kit on pre-prepared cDNA.


All times are GMT -8. The time now is 09:24 AM.

Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2022, vBulletin Solutions, Inc.