home made EZ-Tn5-Nextera adapter complex?

silin284

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Join Date: Jul 2009

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home made EZ-Tn5-Nextera adapter complex?

02-27-2014, 10:28 PM

Hi

Is it possible to mix equal molar of EZ-Tn5 transposase and the Nextera adapter to create a homebrew transposon?

It seems that the transposon in the Nextera kit contain 2 different adapter arms. But the conventional transposase and donor DNA complex should be 1 transposase + 1 DNA. So how did Epicentre prepare their transposon?
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SCIL2019

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Join Date: May 2019

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10-01-2019, 10:02 AM

The transposon showed by Epicentre is mainly for mutagenesis. For NGS applications you load Ez-Tn5 as done for any other (Picelli et al, 2014 for example) using the P5/MErev and P7/MErev. You can do it separately, namely, Tn5 + Px/MErev, or together Tn5 + equimolar mixture of annealed P5/MErev and P7/MErev. In the Transpisition buffer the presence of Mg will induce dimerisation of Tn5-Px/MErevs and it becomes active.
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