This is a test version of Biostars. For the public version, visit https://www.biostars.org.
Is this library suitable for sequencing?

Hello good people,

I am relatively new to NGS library preparation. I prepared these libraries for bulk RNA-seq using the NEBNext Ultra II Directional RNA Library Prep Kit.

The TapeStation profile shows multiple peaks extending into the higher size range. My guess is that I may have over-amplified the library during PCR.

I would greatly appreciate any advice on whether these libraries are suitable for sequencing as they are, or if it would be better to prepare new libraries to ensure better data quality. I was going for 6 Gigabase data in 150PE format.

Thank you in advance for your help.

I have attached the tapestation result for your reference

library ngs rna-seq
• 164 views • link
updated by GenoMax GenoMax • written by Poshan • Poshan

You could use use a procedure like this to clean the libraries: https://knowledge.illumina.com/library-preparation/dna-library-prep/library-preparation-dna-library-prep-reference_material-list/000006157

For future reference, this question is more suitable for SeqAnswers or an experimental biology forum. Don't think your libraries look too bad for sequencing as is. Smaller library fragments cluster preferentially on Illumina FC in general.

• 0 views • link
written by GenoMax GenoMax

0 answers

No answers yet.

Log in to answer this question.

More posts like this