Padlock Capture for DNA Analysis

Padlock capture was performed as previously described^{1 (link),3 (link),15 (link)}. Briefly, each reaction was performed in 20 µl volume containing 1 unit Ampligase (A3210K, Epicentre), 1 unit Phusion High-Fidelity DNA Polymerase (M0530, New England BioLabs), 1 x Phusion High-Fidelity DNA Polymerase buffer, 10 nM dNTP and 1 ng padlock probe. Two microliters of the purified pre-PCR product and 800 ng genomic DNA were used in each reaction. Nicotinamide adenine dinucleotide (NAD+) was provided in each reaction at a final concentration of 0.5 mM.

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Hong R., Chandola U, & Zhang L.F. (2017). Cat-D: a targeted sequencing method for the simultaneous detection of small DNA mutations and large DNA deletions with flexible boundaries. Scientific Reports, 7, 15701.

Publication 2017

Ampligase Phusion High-Fidelity DNA Polymerase

Ampligase Buffer Dna polymerase Genomic Nicotinamide adenine dinucleotide

Corresponding Organization :

Other organizations : Nanyang Technological University

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Variable analysis

independent variables

Reaction volume (20 µl)
Ampligase (1 unit)
Phusion High-Fidelity DNA Polymerase (1 unit)
Phusion High-Fidelity DNA Polymerase buffer (1x)
DNTP (10 nM)
Padlock probe (1 ng)
Purified pre-PCR product (2 µl)
Genomic DNA (800 ng)
Nicotinamide adenine dinucleotide (NAD+) (0.5 mM)

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

positive controls

Not specified

negative controls

Not specified

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