Application of long-distance PCR to restriction site mapping of a cloned DNA fragment on the lambda EMBL3 phage vector.
Long-distance PCR was applied to rapidly map the restriction sites of long inserts cloned on lambda EMBL3 phage vector. The restriction sites of 9 of 15 enzymes were completely assigned in a model experiment within 14 h, including 8h for the PCR amplification. This method was found particularly useful for genomic DNA cloning when the partial sequence of the corresponding cDNA is known.
Machida
M
M
Department of Molecular Biology, National Institute of Bioscience and Human-Technology, Ibaraki, Japan.
Manabe
M
M
Yasukawa
M
M
Jigami
Y
Y
D010582:Bacteriophage lambda / Q000235:genetics*; D001483:Base Sequence; D018076:DNA, Complementary; D004279:DNA, Viral / Q000032:analysis; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D015183:Restriction Mapping
D018076:DNA, Complementary; D004279:DNA, Viral