IGEM:Peking/2007/Count-Conjugation-Notebook/2007-7-24

=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.

Electorphoresis

 * 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.

Ligation

 * 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.