Difference between revisions of "IGEM:Peking/2007/Switch-Notebook/2007-8-14"

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==Colony PCR test of T1T2-(Tet F-lacZ-pLX010)==
 
==Colony PCR test of T1T2-(Tet F-lacZ-pLX010)==
6 colonies are selected for incubation.
+
22 colonies are selected for incubation.
 +
 
 +
==PCR and double digestion of RD1==
 +
===PCR of RD1===
 +
Retract through electropheresis.
 +
 
 +
===Double digestion of RD1===
 +
<pre>
 +
10uL  10*H buffer
 +
  2uL  SpeI
 +
  2uL  XhoI
 +
15uL  RD1
 +
71uL  ddH2O
 +
-------------------
 +
100uL  total
 +
</pre>

Latest revision as of 22:14, 17 August 2007

Ligation of SulA, Rec, SD1-3, SS1-2 with GFP-pLX007

Precipitate for retraction

SulA, Rec, SD1-3, SS1-2, GFP-pLX007 (^^ with XbaI & XhoI)

Ligate with GFP-pLX007

SulA, Rec, SD1-3, SS1-2

Transformed in the evening

Colony PCR test of Prec, SD2-3, SS1-3-(SDY-EGFP-pLX007)

Number of colonies grew up:

       No.
----------
Prec    0
SD2     0
SD3     1
SS1   >50
SS2     2
SS3    14

Only SS1 is right. 4 colonies are select for incubation.

Colony PCR test of T1T2-(Tet F-lacZ-pLX010)

22 colonies are selected for incubation.

PCR and double digestion of RD1

PCR of RD1

Retract through electropheresis.

Double digestion of RD1

 10uL   10*H buffer
  2uL   SpeI
  2uL   XhoI
 15uL   RD1
 71uL   ddH2O
-------------------
100uL   total