SEQanswers - View Single Post - Custom barcoded RNA-seq

link1 · 08-04-2010, 08:46 PM

we tried barcoding the 5'RNA adapter and found significant bias in our small RNA preps. it turns out that RNA Lig I prefers some bases over others. You will need to barcode the 3' adenylated adapter because it utilizes a truncated version of RNA Ligase 2, which does not exhibit as much base preference.
You can design and make these yourself or purchase a set that has already been designed and optimized to eliminate bias. Initially I tried making these myself. Now I purchase the set below. I've done several runs now with multiplexed samples with good results.

http://www.biooscientific.com/Detail...tGenSequencing

08-04-2010, 08:46 PM	#5
link1 Member Location: US Join Date: May 2010 Posts: 21	barcoding small RNAs we tried barcoding the 5'RNA adapter and found significant bias in our small RNA preps. it turns out that RNA Lig I prefers some bases over others. You will need to barcode the 3' adenylated adapter because it utilizes a truncated version of RNA Ligase 2, which does not exhibit as much base preference. You can design and make these yourself or purchase a set that has already been designed and optimized to eliminate bias. Initially I tried making these myself. Now I purchase the set below. I've done several runs now with multiplexed samples with good results. http://www.biooscientific.com/Detail...tGenSequencing