- T-vector -For cloning of dsDNA fragments
with a 3´ single deoxyadenosine overhangs.
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- Description
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- 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.
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- Storage
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Store at -20°C. Avoid exposure to multiple freeze-thaw cycles.
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- Storage buffer
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- 10 mM Tris-HCl pH 7.8, 1 mM EDTA.
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- Storage
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- Store at -20°C.
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- Applications
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- cloning of DNA fragments with 3´-A overhangs
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Quality control
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spectrophotometric control, cloning efficiency, self-ligation assay.
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References
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1. Clark, J.M. (1988) Nucleic Acids Res. 18, 9677-9686. 2. Harrison J.
et al. (1994) Anal. Biochem. 216, 235-236.
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- Catalog #
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- K1
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Pack size
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1 mg/20 ligations
(conc. 50ng/ml)
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