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Old 07-13-2018, 10:28 AM   #1
Zero/1
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Default single-cell cDNA left 90 min at 95C

Briefly, cDNA from single neurons prepared using SMART-Seq4 kit spent 90 minutes at 95C due to human error. Is the cDNA to be discarded or can it still be used for library preparation with Nextera XT and sent to sequencing?

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Old 07-13-2018, 05:53 PM   #2
luc
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Prolonged high heat exposure definitely introduces a lot of damages to DNA. I would expect it to be pretty fragmented by now. I also assume the Nextera transpose would require dsDNA - thus, you would need to add at least another PCR cycle. Perhaps you could use this S1 kit instead?: https://swiftbiosci.com/products/acc...-library-kits/

If the sample can be reproduced, best dump it.

Last edited by luc; 07-13-2018 at 05:55 PM.
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