T-Vector pFB-19T
Code No. FB0015 Size：1 μg Store at － 20℃
Description :
T-Vector pFB-19T is a linearized vector with a single 3’-terminal thymidine at both ends. The T-overhang ends at the cloning site improve the efficiency of ligation of PCR products which contain A-overhangs at 3’-ends.
Vector Map
Components :
T-Vector pFB-19T (50 ng/μl) 20 μl
Experimental Example :
1. Combine the following components in a microcentrifuge tube (total volume 5 μl).
T-Vector pFB-19T 1 μl
PCR product 1μl (0.1 - 0.3 pmol)
ddH2O 3 μl
2. Add one volume (5 μl) of DNA Ligation Kit to the DNA solution and mix thoroughly.
3. Incubate the reactions
4. Add the reaction solution (10 μl) to 100 μl of E. coli competent cells.
Gently flick the microcentrifuge tube to mix and then place it on ice bath for 30 minutes.
5. Heat-shock the cells for 45 seconds in a water bath at 42℃. Immediately return the tube to ice bath for 1 minute.
6. Add 890 μl of SOC medium to the tube and incubate for 1 hour at 37℃ with shaking.
7. Plate 100 μl of transformation culture onto LB/ampicillin plates, then incubate overnight at 37℃.
8. Select the white colonies, and confirm by PCR amplification.
