Introduction
General Information
Storage instruction | Storage at -20 ℃ |
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Advantage |
1.No special equipment required. 2.Time-saving |
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APPLICATION | -- Sticky end ligation -- Blunt end ligation -- TA ligation |
PROTOCOL |
1. Prepare the ligation reaction mixture in a microcentrifuge tube. 5×Quick Ligase Buffer 2 µl Inserta X ng Vectorb ~50 ng T4 DNA Quick Ligase (1 U/µl) 1 µl Sterile distilled Water To 10 µl a. The molar ratio of the insert and vector should be among 3:1 to 8:1. b. For vector with blunt terminal, please perform the dephosphorylation of vector to prevent cyclization. 2. Incubate the reaction mixture 10-15 min at 25℃ for sticky end ligation and 20-40 min at 25℃ for blunt end ligation. 3. The reaction mixture can be used directly to transform. |