M.B. Enzymes GmbH

T-vector -For cloning of dsDNA fragments with a 3´ single deoxyadenosine overhangs.

Description

The T-vector, based on pBluescript KS+ cloning vector, is a ready-to-use linearized plasmid having single 3´-T overhangs on both ends. It is designed to ligate PCR products yielded with thermostable DNA polymerases possesing 3´ terminal deoxynucleotidyl transferase activity that usually results in addition of a single deoxyadenosine to the 3´-end of PCR product. The vector can be used for cloning of PCR fragments generated by DNA polymerases with proof-reading activity, if these products are purified and finally incubated with appropriate DNA polymerase or with TdT and ddATP. For subcloning purposes, sites for restriction endonucleases should be introduced into primers used for amplification by design. Cloned sequences can be transcribed using promoters for both T7 and T3 RNA polymerases flanking a cloning region.

Storage

Store at -20°C. Avoid exposure to multiple freeze-thaw cycles.

Storage buffer

10 mM Tris-HCl pH 7.8, 1 mM EDTA.

Storage

Store at -20°C.

Applications

cloning of DNA fragments with 3´-A overhangs

Quality control

spectrophotometric control, cloning efficiency, self-ligation assay.

References

1. Clark, J.M. (1988) Nucleic Acids Res. 18, 9677-9686. 2. Harrison J. et al. (1994) Anal. Biochem. 216, 235-236.

Catalog #

K1

Pack size

1 mg/20 ligations (conc. 50ng/ml)

T-vector -For cloning of dsDNA fragments with a 3´ single deoxyadenosine overhangs.
Description	The T-vector, based on pBluescript KS+ cloning vector, is a ready-to-use linearized plasmid having single 3´-T overhangs on both ends. It is designed to ligate PCR products yielded with thermostable DNA polymerases possesing 3´ terminal deoxynucleotidyl transferase activity that usually results in addition of a single deoxyadenosine to the 3´-end of PCR product. The vector can be used for cloning of PCR fragments generated by DNA polymerases with proof-reading activity, if these products are purified and finally incubated with appropriate DNA polymerase or with TdT and ddATP. For subcloning purposes, sites for restriction endonucleases should be introduced into primers used for amplification by design. Cloned sequences can be transcribed using promoters for both T7 and T3 RNA polymerases flanking a cloning region.
Storage	Store at -20°C. Avoid exposure to multiple freeze-thaw cycles.
Storage buffer	10 mM Tris-HCl pH 7.8, 1 mM EDTA.
Storage	Store at -20°C.
Applications	cloning of DNA fragments with 3´-A overhangs
Quality control	spectrophotometric control, cloning efficiency, self-ligation assay.
References	1. Clark, J.M. (1988) Nucleic Acids Res. 18, 9677-9686. 2. Harrison J. et al. (1994) Anal. Biochem. 216, 235-236.
Catalog #	K1
Pack size	1 mg/20 ligations (conc. 50ng/ml)