KLENOW FRAGMENT MOLECULAR WEIGHT FULL
KLENOW FRAGMENT MOLECULAR WEIGHT ISO

Diversity, Equity, Inclusion, and Access.Citation, Usage, Privacy Policies, Logo.Biologically Interesting Molecule Reference Dictionary (BIRD).Artificially expanded genetic information system: a new base pair with an alternative hydrogen bonding pattern, Nucleic Acids Res. The use of thymidine analogs to improve the replication of an extra DNA base pair: a synthetic biological system. A third base pair for the polymerase chain reaction: inserting isoC and isoG. A branched DNA signal amplification assay for quantification of nucleic acid targets below 100 molecules/ml. Hybridization properties of the 5-methy-isocytidine/isoguanosine base pair in synthetic oligodeoxynucleotides. Enzymatic incorporation of a new base pair into DNA and RNA. The achievement of high fidelity PCR with the 5-Me-iso-dC:iso-dG base pair opens the door to developing both artificially expanded genetic systems (6) and novel qPCR systems (for example, Promega’s Plexor technology) based on this approach. Using this artificial base pair system (5-Me-iso-dC:iso-dG, dA:dT*, dC:dG) with KF-Taq, the fidelity in PCR was increased to about 98% per amplification cycle. dT*pairs with dA, but not with iso-dG (5). More recently, Sismour and Benner solved this problem by using 2-thio-dT (dT*) in place of dT. The limit in fidelity is chiefly due to the ability of iso-dG’s 1,2 tautomer to mis-pair with dT. In 2004, Johnson and co-workers observed that, by using the Klenow fragment of Taq polymerase (KF-Taq) in PCR, the fidelity of the 5-Me-iso-dC:iso-dG base pair was about 96% per amplification cycle (4).

KLENOW FRAGMENT MOLECULAR WEIGHT FULL

Such optimization is necessary to enable the full development of artificially expanded genetic systems utilizing an expanded genetic code, thereby allowing for the site-specific incorporation of novel functional components (such as unnatural amino acids) into proteins. (b) qPCR and artificially expanded genetic systems: A number of research groups have been working on optimizing PCR amplification on templates containing 5-Me-iso-dC. The limits of detection of the assay were improved 10-fold, from < 500 HIV molecules/mL to < 50 molecules/mL. Use of this strategy resulted in a significant reduction in non-specific hybridization of the above three sequence types to non-target nucleic acid sequences, and thus less amplification of background. For example, Collins and co-workers significantly improved the sensitivity of a branched DNA quantitative hybridization assay for detecting the HIV POL sequence by incorporating ~30% 5-Me-iso-dC and iso-dG into the pre-amplifier, branched DNA (bDNA) amplifier and alkaline phosphate probe sequences used in the assay (3). (a) Molecular recognition: The 5-Me-iso-dC:iso-dG base pair has been incorporated into hybridization assays to enhance probe-target specificity and reduce spurious hybridization to non-target sequences.

The combination of 5-Me-iso-dC’s high selectivity for iso-dG, and the resulting base pair’s high thermodynamic stability, make this modified base particular attractive in the following applications:

KLENOW FRAGMENT MOLECULAR WEIGHT ISO

Furthermore, since iso dC and 5-Me-iso-dC does not pair with dG, iso dC and 5-Me-iso-dC:iso-dG can function as a stable unnatural base pair that can be used to expand the genetic code. Substitution of a 5-me iso-dC:iso-dG base pair for a C:G pair increases the Tm of the resulting duplex by ~2degC per base pair substitution (1,2). Iso dC and 5-methyl iso-deoxycytosine (5-Me-iso-dC) forms a Watson-Crick base pair with iso-dG, but has a different type of hydrogen bonding pattern than those observed for the natural base pairs A:T and C:G.

Klenow fragment molecular weight