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Old 10-07-2019, 03:34 PM   #1
chilipep
Junior Member
 
Location: NYC

Join Date: Oct 2019
Posts: 2
Default n00b Smart-seq2 questions

Hi there, I'm new to Smart-seq and have a couple questions:

1. Can the dT(30)VN primer be substituted with dT(20)VN, dT(23)VN, etc.?
2. How does Superscript IV compare to SSII or SSIII?
3. Is KAPA HiFi HotStart still the best option for preamplification?

Finally a very basic question...
If I am doing targeted amplification of a single gene, can I just use my custom primers for PCR after cDNA preamp and skip all the tagmentation steps? I just want to see whether a heterozygous mouse is picking one allele and silencing the other. I don't need the high resolution of NGS, but instead will TOPO clone these single amplicons and pick enough representative clones to decide whether the alleles are being equally expressed or not.

Thanks for your patience with my n00b self!
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