IGEM:Peking/2007/Count-Conjugation-Notebook/2007-7-24: Difference between revisions

Revision as of 11:08, 24 July 2007

pls see my suggestion @notebook 2007-7-23 -by QuMingZhi i will be back @29/July ,add oil ,all!

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.

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 pls see my suggestion @notebook 2007-7-23 -by QuMingZhi
 i will be back @29/July ,add oil ,all!
+===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.
+[[Image:crRNA and R0040 test.jpg]]
+===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, overnight.