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

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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.