[An effective method for site-directed mutagenesis in plasmids and cloning single-stranded DNA fragments].
A three primer variant of the earlier devised oligonucleotide-directed mutagenesis in plasmids is described, useful also for the fast cloning of single-stranded DNA products of the asymmetric polymerase chain reaction (PCR). Using this method for plasmid pHD-001-14-11, a 59 b. p. deletion and a 7 b. p. insertion were simultaneously introduced at 81% frequency, and the PCR-copied phage fd transcription terminator (26 b. p.) was inserted with the yield of 67%.
Drutsa
V L
VL
Kaberdin
V R
VR
Koroleva
O N
ON
Shilov
I A
IA
D001345:Autoradiography; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005814:Genes, Viral; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D016133:Polymerase Chain Reaction; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic
D004277:DNA, Single-Stranded
Effektivnyĭ metod napravlennogo vvedeniia mutatsiĭ v plazmidy i klonirovaniia odnotiazhevykh fragmentov DNA.