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Revision as of 02:38, 13 August 2012 (view source) Nili (Talk | contribs) ← Older edit		Revision as of 02:40, 13 August 2012 (view source) Nili (Talk | contribs) Newer edit →
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	*Primer R(10μM) 4μL		*Primer R(10μM) 4μL
	*Template(E.coli DH5α)		*Template(E.coli DH5α)
-	**'''Reflection:You should take less colony.'''	+	**'''Reflection:Took colony too many.You should take less colony.'''
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	==8/11==		==8/11==

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Revision as of 02:40, 13 August 2012

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During the creation

Contents 1 8/9 1.1 Colony PCR 2 8/11 2.1 The purified DNA1,2 2.2 PCR Product 1,3,9

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.

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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

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