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Old 11-23-2016, 04:46 AM   #1
vtorda
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Location: Szeged, Hungary

Join Date: Oct 2016
Posts: 4
Default SmartSeq cDNA validation with TapeStation

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!!
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Old 11-23-2016, 11:50 AM   #2
nucacidhunter
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Location: Iran

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Default

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 at 11:52 AM.
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cdna synthesis, single cell analysis, smartseq2

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