Enzymatics特约代理商|Y9140-LC-L End

Cohesive End Ligation

(equimolar or 2 or 3 X molar concentration of vector) 　　water added to 18 µl 2) Heat the mixture at 45 ℃ for 5 min. to melt any annealed cohesive ends. 3) Cool on ice. 　　Add ligase buffer ( ATP) 2 µl 　　ligase 0.5 µl

Cohesive End Ligation

1) The ligation mixture contains the following: vector (~100 ng)　　insert (equimolar or 2 or 3 X molar concentration of vector)　　water added to 18 µl 2) Heat the mixture at 45 ℃ for 5 min. to melt any annealed cohesive ends. 3) Cool on ice. 　　Add

BAC End-Sequencing

. For example: If DNA is OK for end-sequencing, then prepare 2 reactions for each BAC using T7 and SP6 primers (18mers). 1 reaction:  22.0 µl DNA 16.0 µl reaction mix (ABI/PE #402122) 2.0 µl 25 µM T7 or SP6 (T7 5'-ATTTAGGTGACACTATAG-3') (SP