Team:Tsinghua/Notebook/23 August 2010

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(Difference between revisions)
(New page: == Module I, group 2c == Run the same PCR program as yesterday and purify the product with gel purification kit. ---- Digest eGFP with XbaI and EcoRI, Ap is uesd to phosphorylate the 5' e...)
(→Module I, group 2c)
 
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== Module I, group 2c ==
== Module I, group 2c ==
-
Run the same PCR program as yesterday and purify the product with gel purification kit.
 
----
----
-
Digest eGFP with XbaI and EcoRI, Ap is uesd to phosphorylate the 5' end of digestion result.
+
Digest eGFP with XbaI and EcoRI.
digestion system
digestion system
  eGFP          3ul
  eGFP          3ul
Line 11: Line 10:
  H2O            14ul
  H2O            14ul
----
----
-
Digest mCherry fragment with Dra III,Ap is uesd to phosphorylate the 5' end of digestion result.
+
Digest mCherry fragment with Dra III.
digestion system
digestion system
-
  mCherry        4ul
+
  mCherry        3ul
  FD Buffer      2ul
  FD Buffer      2ul
  Dra III        1ul
  Dra III        1ul
  H2O            14ul
  H2O            14ul
----
----
-
Digest Kan fragment with Dra III and SpeI,Ap is uesd to phosphorylate the 5' end of digestion result.
+
Digest Kan fragment with Dra III and SpeI.
digestion system
digestion system
-
  Kan           4ul
+
  Kan           3ul
  FD Buffer      2ul
  FD Buffer      2ul
  Dra III        0.5ul
  Dra III        0.5ul
-
  AP            0.5ul
+
  SpeI          0.5ul
-
  H2O            13ul
+
  H2O            14ul
----
----
-
Digest plasmid PSB1C13 with XbaI and SpeI, AP is used to phosphorylate the 5' end of digestion result.
+
Digest Chlr fragment with EcoRI and PstI.
 +
digestion system
 +
Kan            3ul
 +
FD Buffer      2ul
 +
XbaI          0.5ul
 +
Pst I          0.5ul
 +
H2O            14ul
 +
----
 +
 
 +
Digest plasmid PSB1C13 with XbaI and SpeI, AP is used to dephosphorylate the 5' end of digestion result.
digestion system
digestion system
  PSB1C13        4ul
  PSB1C13        4ul

Latest revision as of 20:20, 26 October 2010

Module I, group 2c

Digest eGFP with XbaI and EcoRI. digestion system 

eGFP           3ul
FD Buffer      2ul
XbaI           0.5ul
Dra III        0.5ul
H2O            14ul

Digest mCherry fragment with Dra III. digestion system 

mCherry        3ul
FD Buffer      2ul
Dra III        1ul
H2O            14ul

Digest Kan fragment with Dra III and SpeI. digestion system 

Kan            3ul
FD Buffer      2ul
Dra III        0.5ul
SpeI           0.5ul
H2O            14ul

Digest Chlr fragment with EcoRI and PstI. digestion system 

Kan            3ul
FD Buffer      2ul
XbaI           0.5ul
Pst I          0.5ul
H2O            14ul

Digest plasmid PSB1C13 with XbaI and SpeI, AP is used to dephosphorylate the 5' end of digestion result. digestion system 

PSB1C13        4ul
FD Buffer      2ul
XbaI           0.5ul
SpeI           0.5ul
AP             0.5ul
H2O            12.5ul

12h after digestion, the products are purified with kit.

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