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Old 02-06-2019, 03:25 AM   #1
Fullmetal
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Location: Europe

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Default Mechanism preventing unspecific Binding of Adapter post A overhang introduction (10x)

Hello All,

I'am designing an scRNA seq experiment using 10x genomics, when I look at their chemistry, I had a few questions that remained unanswered, I hope someone here could help me out.

In the cDNA amplification process for library prep, there is a step for introduction of an A overhang after fragmentation to facilitate adapter binding, but the overhang is introduced on both 3' ends of the cDNA fragment, but the adapter ligation happens only at one end and the correct end at that.

Does anyone know if there are any enzymes or mechanisms that facilitate this process? or if you understand this mechanism better could you please enlighten me a little about it.

Thanks
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