MoFavor™ Cloning Kit Code  BK0030-0020 Classification  CLONING Size  20rxns Order  Print  PRODUCT DETAIL Description   The MoFavor™ Cloning Kit is convenient for the cloning of PCR products generated using Taq and other polymerase that do not possess 3 '- 5' exonuclease activity. this kit offers high cloning effeciency with 90% of the recombinants containing inserts.   Features   The 3'-protruding nucleotides of the vector are stabilized to promote transformation effciency.. Easy blue/ white colony screening. M13 forward and reverse primer sites for DNA sequencing. The kit contains Ligation Master Mix, linear form cloning vector and control insert DNA.   Feature Position (bp) Multiple cloning site (MCS) 397-463 IacZ -αpeptide 486-149 pUC origin 882 Ampicillin resistance gene 2501-1644 Primer binding sites:      M13 forward primer 359-375    M13 reverse primer 496-480 Kit Components   MoFavorTM Cloning Kit contains three components: Cloning Vector, Ligation premix and Control Insert DNA. The Cloning Kit is supplied in a ready-to-use linear form for direct ligation of PCR products. The Ligation Master Mix is a cocktail containing ligase, buffer and cofactors to support ligation. The control insert DNA is a 847 bp fragment. This provides a positive control for he ligation reaction.   Procedure   Kit is designed for a 10 μl reaction volume containing 5 μl of the Ligation Master Mix plus varying volumes of vector, insert and water (if needed). Prepare ligation reaction and incubate for several hours or overnight at 4℃. Heat the ligation mixture in 50 ℃ for 10 minutes before transformation. Transform competent cells with 1-2 μl of ligation mixture. Select using on ampicillin-containing medium.   Notes The Ligation Master Mix contains ATP and should be aliquoted as frequentfreezing and thawing causes ATP hydrolysis.. Mix the thawed ligation buffer well before use. The ligation mixture should be homogenous as well. The number of transformant is proportional to theduration of ligation. Overnight ligation at 4℃ is suggested.