High quality Bacterial Artificial Chromosome (BAC) libraries form an important basis in KeyGene's genetic and genomic research. KeyGene has constructed a wide range of BAC libraries for own use and for external parties.
How it works
KeyGene's BAC library construction protocol consists of the following steps:
isolation of HMW-DNA according to Liu et.al.;
partial digestion of the plugs with HindIII / EcoRI / BamHI;
first and second sizing steps by pulsed field electrophoresis (CHEF);
electroelution and quantification of HMW-DNA for ligation;
ligation in pIndigoBAC-536 vector;
electroporation of ligated HMW-DNA into E. coli DH10B cells;
NotI sizing of ligated BAC clones;
large electroporation of top ligations ( > 120 kb);
picking of the BAC clones in 384 well plates - Optional : Making of high density filters;
quality control of the BAC library - insert sizing of NotI digests after picking.
technologies 
