T&A Cloning Kit

Description

The T&A Vector System is convenient for cloning PCR product generated using thermostable DNA polymerases, such as RealTaq Polymerase, which add a single terminal 3'-dA nucleotide overhang. Ligation can be achieved in as little as 5 minutes. Following ligation the mixture may be used directly to transform HIT™ or other competent cells or purified to achieve a higher efficiency of transformation.

Applications and Benefits

Accepts terminal 3'-dA nucleotide overhang PCR products

Transform ligation product (purified/unpurified) into HIT™ cells

LacZ Complementation for Blue/White screening

Ampicillin marker for antibiotic selection

Universal primer for easy transformation screening

Ligation Conditions	User Sample	Positive Control
Ligation Buffer A	1µl	1µl
Ligation Buffer B	1µl	1µl
T&A cloning vector	2µl	2µl
PCR product	Xµl	********
T4 DNA ligase	1µl	1µl
Control DNA	********	3µl

Add deionized water to a final volume of 10 μl Incubate the reactions for 5 to 15 min at 220C.

Cat. No. RC001 (20 rxn)
T&A Cloning Vector (40 ul, 25 ng/ul)
Control Insert DNA (5ng/ul) (10 ul)
T4 DNA Ligase (3 Ul/ul) (20 ul)
T4 DNA Ligase Buffer A (100 ul)
T4 DNA Ligase Buffer B (100 ul)
Forward primer (M13-F) (10 uM) (50 ul)
Reverse primer (M13-R) (10 uM) (50 ul)

Cat. No. RC013
T&A (25ng) ul Cloning Vector (40 ul)