View Single Post
Old 12-21-2015, 07:13 PM   #176
kobeho24
Member
 
Location: HKUST, Hong Kong

Join Date: Apr 2015
Posts: 32
Thumbs up

Quote:
Originally Posted by Simone78 View Post
Hi Gary,
yes, I saw the paper, thanks! I didnīt know that the TGIRT can do strand switch on DNA as well (this was not mentioned when I contacted the Lambowitz lab). When I first did my tests some time ago I used tot RNA. Unfortunately, it didnīt work at single cell level (pg) and I just dropped it. therefore, I never sequenced anything and never tried on a real cell. I do have a protocol that worked with 100 ng (!) RNA but after the publication of this new article, I want to try again using some modifications and see if I can get it to work with less RNA. So I canīt really share it right now, sorry!
Best,
Simone
Hi Simone,
Sounds really great that you do have a protocol. What I wonder is RT condition for non-fragmented RNA. Just incubate the RNA with TGIRT at proper temperature for 15 min? Looking forward to your successful modified protocol.

Gary
kobeho24 is offline   Reply With Quote