NGS dA-Tailing Module

The NGS dA-Tailing Module is used to efficiently incorporate a non-templated dAMP on the 3´ end of a blunt DNA fragment. 3´-dA DNA tailing prevents concadamer formation during subsequent ligation steps. DNA tailed with the NGS dA-Tailing module may be ligated to adaptors or cloning vectors with complementary dT overhangs. The NGS dA-Tailing Module is provided as a master mix to maximize efficiency and convenience in DNA sample preparation workflows.

The module is optimized for use with the NGS Quick Ligation Module (Cat #E114-1, E114-2), and is part of the NGS DNA library prep workflow for Illumina sequencing.

Applications

• dA-Tailing of fragmented DNA.
• DNA sample preparation.

Protocol 

1. Assemble the following reaction in a microcentrifuge tube on ice:

     End Repaired, Blunt DNA                               1-5 µg
     dA-Tailing Reaction Buffer (10×)                     10 µL
     Klenow Fragment (3'→5' exo^–)                        3 µL
     Nuclease-free water                                        to 50 µL
    Total volume                                                    50 µL

2. Mix gently and spin down for a few seconds.

3. Incubate at 37°C for 30 min.

4. Purify DNA Sample on one spin column.