A new approach for point mutation detection based on a ligase chain reaction.
A new method for the identification of point mutations is proposed. The method is based on ligase chain reaction (LCR) and it includes a procedure for correction of ligation by Cleavase. Reaction products are detected by a colorimetric method after adsorption of the resulting DNA duplexes to the solid phase. One strand of LCR products carries biotin to be bound on a streptavidin-coated microwell. Another strand contains a single-stranded region that is to be coupled with an oligonucleotide carrying a substrate for colorimetric detection. The suggested method has two advantages: (i) use of Cleavase increases the accuracy of ligation and (ii) a template independent ligation does not occur in LCR due to a special design of primers.
Demchinskaya
A V
AV
Institute of Agricultural Biotechnology, Timiryazevskaya ul. 42, 127550, Moscow, Russia.
Shilov
I A
IA
Karyagina
A S
AS
Lunin
V G
VG
Sergienko
O V
OV
Voronina
O L
OL
Leiser
M
M
Plobner
L
L
D001483:Base Sequence; D001710:Biotin / Q000378:metabolism; D011088:DNA Ligases / Q000378:metabolism*; D004252:DNA Mutational Analysis; D004591:Electrophoresis, Polyacrylamide Gel; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D009154:Mutation; D009841:Oligonucleotides / Q000737:chemistry; D017354:Point Mutation; D012689:Sequence Homology, Nucleic Acid
D009841:Oligonucleotides; D001710:Biotin; D004706:Endodeoxyribonucleases; C118972:cleavase; D011088:DNA Ligases