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-   -   TET,EBT, Tween-20 (http://seqanswers.com/forums/showthread.php?t=94675)

SofiaGranja 06-03-2020 08:44 AM

TET,EBT, Tween-20
 
Hi everyone,

I have two questions:

There are a considerable number of DNA extraction protocols that indicate to elute in TET (10 mM Tris – HCl + 1 mM EDTA + 0.05% Tween 20). However, the presence of EDTA can inhibit future reactions such as library build. Any idea why the protocols are not standardised to always use EBT in the elution?

Regarding to Tween 20, it is a non-ionic detergent that reduces adsorption of DNA to plastics and improves pipetting accuracy. However, if we use low binding tubes, adding tween 20 to TE or EB, it is not redundant or even useless to use it? (since the technology that these optimized tubes present nearly 100% recovery of DNA / RNA molecules).

Can't wait to hear your thoughts on this.

Thank you,
Sofia :)

luc 06-05-2020 05:20 PM

I have never seen any problems with DNA samples in EBT or EB buffer.
This tiny amount of detergent does not seem to cause problems.
I agree with avoiding EDTA buffers;at least at 1 mM concentrations. 0.1 mM EDTA (as in TLE buffer) also does not seem to cause problems.


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