Team:Kyoto/GSDD/Notebook/0921-0927



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To Do

 * 1) Colony PCR
 * Insert check
 * 1) Miniprep
 * Restriction enzyme digestion
 * Gel extraction and PCR purification
 * Ligation
 * Transformation

PCR

 * ①and ②and ③ are inserted properly.

Miniprep



 * Gel extraction:4(1)a E-S,1-0-2 X-P
 * PCR purification:4(2) E-X

To Do

 * 1) Miniprep 3(1)a1 4(4)a1 11①
 * Restriction enzyme digestion
 * Gel extraction and PCR purification
 * Ligation
 * MPR

PCR
Gel extraction and PCR purification Sample 4(4)E-X, 3(1)E-S

To Do

 * 1) Phosphorylation
 * 2) Miniprep
 * Restriction enzyme digestion
 * BAP Gel extraction and PCR purification
 * Ligation

electrophoresis
11-1 can be cut by stuI. 4(5) can not be inserted in Byp 561 because 4(5) has PstI site.

Dephosphorylation



 * Gel extraction.


 * PCR purification

To Do

 * 1) Restriction enzyme digestion (Stu1)
 * Transformation
 * 1) Transformation
 * 2) Miniprep
 * Restriction enzyme digestion
 * Ligation
 * Transformation
 * 1) Ligation
 * transformation

To Do

 * 1) Culture 1M 2M(1) 11 (1)
 * Restriction enzyme digestion
 * Insert check
 * 1) Electorophoresis 4(5)a,b,c
 * 2) Colony PCR

Electrophoresis
1M contains product of MPR with 200bp. 2M ,at first, contains product of MPR with 1kbp. With transformed, it changed to with 600bp.