A fast and efficient method for isolation of the BAC end.
As a new developmental vector system, the bacterial artificial chromosome (BAC) has been used widely in constructing genomic libraries and in generating transgenic animals. Isolation of the BAC insert end is useful to analyze the BAC clone. Here, we describe a fast and efficient method to obtain the BAC end by ligating the BAC fragments digested with Not I and another selected restriction enzyme into universal cloning vector, followed by determining the correct clones with HindIII digestion. Further DNA sequencing analysis verified the results mentioned above.
Huang
Y
Y
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, 5 Dong Dan San Tiao, Beijing, 100005, China.
Liu
D P
DP
Wu
M
M
Liang
C C
CC
D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D015247:Deoxyribonuclease HindIII / Q000494:pharmacology; D015252:Deoxyribonucleases, Type II Site-Specific / Q000494:pharmacology; D008967:Molecular Biology / Q000379:methods*; D010957:Plasmids / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013997:Time Factors
D004262:DNA Restriction Enzymes; D015247:Deoxyribonuclease HindIII; D015252:Deoxyribonucleases, Type II Site-Specific; C038565:GCGGCCGC-specific type II deoxyribonucleases