ACEQuick ligation Kit contain 5x Quick ligation buffer and T4 DNA ligase that catalyzes the formation of a phosphodiester bond between adjacent 5'-phosphate and 3'-hydroxyl termini in double-strand DNA or RNA. This kit is suitable for sticky end ligation, blunt end DNA ligation and TA ligation. Buffer with optimized ligation enhancer make the reaction more efficient and convenient. Ligation can be completed in 10 min at 25℃ for sticky end ligation and 20 min at 25℃ for blunt end DNA ligation . The product can be used directly to transform many chemically competent cells.
General Information
Storage instruction
Storage at -20 ℃
Advantage
1.No special equipment required.
Ligation can be completed at 25℃ without equipment that has cooler for 16℃ setup.
2.Time-saving
Ligation can be completed in 10 min for sticky end ligation and 20 min for blunt end DNA ligation .
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.
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