Team:KAIT Japan/Notebook

From 2012.igem.org

(Difference between revisions)
Line 14: Line 14:
During the creation<br>
During the creation<br>
-
 
+
<big>Creating parts of Tar methylation region.<big>
==8/9==
==8/9==
===Colony PCR===
===Colony PCR===
Line 25: Line 25:
*Template(E.coli DH5α)
*Template(E.coli DH5α)
→'''Reflection:Took colony too many.You should take less colony.'''
→'''Reflection:Took colony too many.You should take less colony.'''
 +
----
----
==8/11==
==8/11==

Revision as of 02:53, 13 August 2012

KAIT_Japan_Rogo2mini.png

Home Team Project Parts Attributions Notebook Safety Sitemap


During the creation
Creating parts of Tar methylation region.

Contents

8/9

Colony PCR

Reagent

  • TaKaRa Ex Taq(5units/μL) 0.5μL
  • 10×Ex Taq buffer 10μL
  • dNTP Mixture(2.5Meach) 8μL
  • Primer F(10μM) 4μL
  • Primer R(10μM) 4μL
  • Template(E.coli DH5α)

Reflection:Took colony too many.You should take less colony.


8/11

The purified DNA1,2

  1. Electrophoresis
    • Marker:pigment(buffer) 1μL,DNA molecule 2μL,TE buffer 3μL
    • Sample:pigment(buffer) 1μL,sample 5μL
    • Gel concentration:1.2%,Migration time:30minutes
  2. Storage

PCR Product 1,3,9

  1. Electrophoresis
    • The gel check and cut
  2. DNA purification
  3. Confirmation of electrophoresis
  4. PCR
    • 50cycle
  5. Storage