View Single Post
Old 07-30-2010, 07:20 AM   #1
nasobema
Member
 
Location: Germany

Join Date: Jul 2010
Posts: 14
Default Custom barcoded RNA-seq

Hi,

we are planning on setting up our own custom method for RNA-sequencing with barcodes on an Illimina GAII. I've already found some hints in this forum and elsewhere but still have some questions:

- My plan is to the use Illumina Small RNA adapters and corresponding PCR primers (like they are displayed at http://seqanswers.com/forums/showthr...pters+sequence) with and additional barcode of 4-6 nucleotides added to the 5' RNA Adapter.
Does anyone have experience with that and wants to share it?

- After RT, do I need a second strand synthesis at all? If I get it right, a PCR on the ss-cDNA should do the job and generate a ready-to-use cDNA library with adapters and all.

Any help is highly wellcome.
nasobema is offline   Reply With Quote