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Old 03-20-2017, 08:32 AM   #1
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Location: Chicago

Join Date: Mar 2017
Posts: 6
Default 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 dNTPs present in the catching buffer that you sort cells into? Couldn't they just be added with the RTase after the initial binding of the OligoDT to the polyA tail?

2) What are the drawbacks of having OligoDTs or dNTPs in excess to what is written in the original protocol? I am worried about the concentrations not being enough for a highly transcriptional cell.

Thanks in advance for the help!
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