Key & Lock by Yu Tao and Zheng Qinsi
Purification of the Digested Fragment and Vector
- Fragment: crRNA(J23078); Vector: R0040.
- Use PCR Purification Kit.
- The volume of purified fragment and vector: each 30uL, respectively.
- To check the yield of the purified fragment and vector.
- 2% agarose.(For the ~70bp crRNA)
- Each sample: 2uL; with 1uL 10*loading buffer and 1uL 200* DNA dye added, respectively.
- From left to right: crRNA_PstI+XbaI, R0040_PstI+SpeI, R0040 plasmid, Marker I.
- Ligate the crRNA fragment and R0040 vector.
- crRNA 2uL, R0040 4uL, totally 10uL system.
- system without crRNA added as a negative control.
- 4 centigrade degree, overday.
- Then transform them.