诺唯赞试剂说明书 T4 DNA Ligase.pdfVIP

Sterile distilled water Up to 10 μl 10x Ligase Buffer 1 μl T4 DNA Ligase Insert a 0.3 pmol Vector b 0.3 pmol Catalog # : GM40501 T4 DNA Ligase (400 U / μl) 1 μl a. The molar ratio of the insert and vector should be among 3:1 to 10:1. b. For vector with blunt terminal, please perform the dephosphorylation of Introduction vector to prevent cyclization. T4 DNA ligase catalyzes the formation of a phosphodiester 2. Incubate the reaction mixture at 16°C overnight. bond between 5’ terminal phosphate and adjacent 3’ hydroxyl termini in duplex DNA, RNA or DNA/RNA hybrids. 3. Transformation This enzyme will join blunt end and cohesive end termini but it cannot repair single stranded nicks. This product is 1) Take the competent cells out of the -80°C refrigerator, applicable to 3 end labeling of RNA, cyclization of RNA and and place the competent cells immediately in an ice water DNA oligonucleotide and cDNA cloning. bath. 2) Add the DNA into 100 μl of competent cel