DNA Quick Ligation Kit
Lot # Check on the product label
Size: 60 T
Storage: Store at -20°C.
--------------------------------------------------------------------------------------------------------------
Price: $130.00
Catalog# BPC1099-2
Lot # Check on the product label
Size: 60 T
Storage: Store at -20°C.
Application: This kit can quickly complete the ligation of DNA fragments with sticky or blunt ends at room temperature (22C) for 10 minutes. The ligation product can be converted directly. It can be used for DNA fragment cloning, library construction, TA cloning, Linker connection, etc.
|
Components |
Size |
|
2 xQuick Ligation Buffer |
300 μl |
|
T4 DNA ligase, 5 U/μl |
300 U |
Protocol
1. Recommended reaction system
|
2 xQuick Ligation Buffer |
5 μl |
|
Vector DNA (30-60ng) |
μl |
|
Insert DNA (3 times the molar amount of vector, insert and vector molar ratio of 3:1) |
μl |
|
sterile water |
μl |
|
T4 DNA Ligase |
1 μl(5 Weiss Units/ml) |
Final Volume 10 μl
Generally, T4 DNA Ligase is added at the end, mixed well, and the liquid can be collected by instant centrifugation to the bottom of the tube.
2. 22°C connection for 10 minutes (usually can be done in a PCR instrument).
Usually 5-10 minutes is recommended for 22°C ligation, and the ligation of >3kb long fragments can be extended to 30 minutes. It is not recommended to exceed 30 minutes, as it may reduce the number of transformants.
3. Cool on ice, then directly transform or store at -20°C.
If stored at -20°C, it should be redissolved on ice before transformation, and the transformation should be carried out after thorough mixing.
Instruction:
1. Do not heat inactivate T4 DNA ligase before transformation, otherwise the transformation efficiency will be reduced.
2. If the volume of DNA fragments is greater than 4 l, 2 xQuick Ligation Buffer can be added, and the total reaction volume can be increased to 20 l accordingly, so that larger volumes of DNA fragments can be added to the ligation system.
