Scarless assembly of unphosphorylated DNA fragments with a simplified DATEL method.

Abstract:

:Efficient assembly of multiple DNA fragments is a pivotal technology for synthetic biology. A scarless and sequence-independent DNA assembly method (DATEL) using thermal exonucleases has been developed recently. Here, we present a simplified DATEL (sDATEL) for efficient assembly of unphosphorylated DNA fragments with low cost. The sDATEL method is only dependent on Taq DNA polymerase and Taq DNA ligase. After optimizing the committed parameters of the reaction system such as pH and the concentration of Mg2+ and NAD+, the assembly efficiency was increased by 32-fold. To further improve the assembly capacity, the number of thermal cycles was optimized, resulting in successful assembly 4 unphosphorylated DNA fragments with an accuracy of 75%. sDATEL could be a desirable method for routine manual and automated assembly.

journal_name

Bioengineered

journal_title

Bioengineered

authors

Ding W,Weng H,Jin P,Du G,Chen J,Kang Z

doi

10.1080/21655979.2017.1308986

subject

Has Abstract

pub_date

2017-05-04 00:00:00

pages

296-301

issue

3

eissn

2165-5979

issn

2165-5987

journal_volume

8

pub_type

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