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Forum: Sample Prep / Library Generation 07-21-2017, 07:36 AM
Replies: 6
Views: 2,213
Posted By PrimerBuffalo
Yes, that is incredibly helpful thank you! I...

Yes, that is incredibly helpful thank you!

I will definitely reduce my input concentration and increase my number of cycles. I have a colleague who uses 12 cycles for an input of 0.5 ng and I...
Forum: Sample Prep / Library Generation 07-20-2017, 08:36 AM
Replies: 6
Views: 2,213
Posted By PrimerBuffalo
Hi Simone78, I am currently generating...

Hi Simone78,

I am currently generating indexed libraries from my samples and have run into a problem:

I used 1ng of cDNA and ran the indexing PCR for 7 cycles. I did this because I thought...
Forum: Sample Prep / Library Generation 05-11-2017, 01:28 PM
Replies: 1
Views: 2,994
Posted By PrimerBuffalo
UMI in Smart-seq2 protocol

Hi All,

Has anyone had any experience trying to add UMI to the smart-seq2 protocol? I've done some searching, but I haven't found an account of anyone doing so. I would like to know if it is even...
Forum: Sample Prep / Library Generation 04-14-2017, 09:15 AM
Replies: 6
Views: 2,213
Posted By PrimerBuffalo
Thank you so much! That is very reassuring!

Thank you so much! That is very reassuring!
Forum: Sample Prep / Library Generation 04-11-2017, 09:23 AM
Replies: 6
Views: 2,213
Posted By PrimerBuffalo
Variance in cDNA amplification

Hello,

I recently generated cDNA from 100 T-cells (from the same source) using the Smart-Seq2 protocol from Sandberg et al. After the pre-amplification step (all wells were pre-amplified for 18...
Forum: Sample Prep / Library Generation 03-20-2017, 08:32 AM
Replies: 3
Views: 2,776
Posted By PrimerBuffalo
Smart-seq2 sample prep questions

Hi All,

I have a couple questions about the Smart-seq2 method for amplifying single cells (Sandberg 2014), and I hope someone with experience in this method might have some answers:

1) Why are...
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