Reducing nontemplated 3' nucleotide addition to polynucleotide t

Details Reducing nontemplated 3' nucleotide addition to polynucleotide t Reducing nontemplated 3' nucleotide addition to polynucleotide t

1999-08-10

2000-07-18

Sisson, Bradley L.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435 6, 435 912, 536 231, 536 2372, 536 243, 536 2433, C12P 1934, C12Q 168, C07H 2102, C07H 2104

Patent

active

060905905

ABSTRACT:
Non-template 3' nucleotide addition to a transcript is reduced by transcribing a transcript from a template comprising an ultimate and/or penultimate 5' ribose having a C'2 substituent such as methoxy, which reduces non-template 3' nucleotide addition to the transcript. The methods are shown to be applicable to a wide variety of polymerases, including Taq, T7 RNA polymerase, etc.

REFERENCES:
Moran et al., Non-hydrogen bonding `terminator` nucleosides increase the 3'-end homogeneity of enzymatic RNA and DNA synthesis. Nucl. Acids. Res. 24, 2044-2052, 1996.
Tunitskaya et al., Substrate properties of C' -methyl derivatives in T7 RNA polymerase reactions. Evidence of N-type NTP conformation. FEBS Lett. 400, 263-266, 1997.

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