Autor: |
Kimoto, Michiko, Yokoyama, Shigeyuki, Hirao, Ichiro |
Předmět: |
|
Zdroj: |
Biotechnology Letters; Jun2004, Vol. 26 Issue 12, p999-1005, 7p |
Abstrakt: |
A method to determine the steady-state kinetic parameters of single-nucleotide insertion in replication was developed using an automated DNA sequencer. The insertion of nucleoside 5′-triphosphates into a 6-carboxyfluorescein-labeled primer by DNA polymerase was quantified from the band pattern on a gel using GeneScan software. The parameters determined by this method were consistent with those obtained by the conventional radioisotope-labeling method. This non-radioactive, fluorescent-based method is rapid and can handle a large number of samples to assess cognate or non-cognate base pair formation between natural or unnatural bases in replication. [ABSTRACT FROM AUTHOR] |
Databáze: |
Complementary Index |
Externí odkaz: |
|