SmartSeq cDNA validation with TapeStation

vtorda

Junior Member

Join Date: Oct 2016

Posts: 4
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#1

SmartSeq cDNA validation with TapeStation

11-23-2016, 05:46 AM

Hi All,

I have RNA extractions from fungal fruiting body tissue, produced by cryo-sectioning-->LCM-->PicoPure RNA extraction kit.
I have started to use SmartSeq V4 to produce cDNA. After the amplification of the cDNA we checked the cDNA profile by TapeStation.

But the profile doesn’t look like as SmartSeq manual suggests: we have a peak around 400bp instead of 2500bp.
Anybody has experienced cDNA profile different from the profile in SmartSeq manual?
The company who will do sequencing said that this profile is similar what they usually see on cDNA produced by SmartSeqV4.
What is your opinion?

Thank you all!!
Attached Files

smartseq_amplified_cDNA_TapeStation.jpg (30.9 KB, 299 views)
Tags: cdna synthesis, single cell analysis, smartseq2
nucacidhunter

Jafar Jabbari

Join Date: Jan 2013

Posts: 1250
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#2

11-23-2016, 12:50 PM

It looks fine. I would clean it with 0.6x beads before shearing and library prep for RNA-Seq.

Last edited by nucacidhunter; 11-23-2016, 12:52 PM.
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SmartSeq cDNA validation with TapeStation

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