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Revision as of 06:23, 27 August 2010 (view source) Bambi75 (Talk | contribs) (New page: {{:Team:Tokyo_Metropolitan/Header}} ==2010/08/26(Bambi75)== ==Make Plates== ===member=== NEX , Bambi75 and watachin ===Materials=== *RO water 200ml *LB Broth 4g *Cam(50μg/L) 10ml =...) ← Older edit		Revision as of 05:41, 8 September 2010 (view source) NEX (Talk | contribs) (→PCR) Newer edit →
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	**10℃　∞		**10℃　∞
	※30cycle ☆ to ★.		※30cycle ☆ to ★.
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-	==PCR==
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-	===member===
-	NEX , Bambi75 and watachin
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-	===Materials===
-	*sterilized water 71μl
-	*Ex taq buffer 10μl
-	*dNTP mix 8μl
-	*Ex taq 1μl
-	*primer(bcsC sense/antisense) 5μl each
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-	===Procedure===
-	①mix materials
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-	②divide ① into 2 tubes.
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-	③elongation
-	**95℃ 3min
-	**96℃ 1min☆
-	**55℃ 7min
-	**72℃ 1min★
-	**10℃ ∞
-	※30 cycle ☆to★.

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Revision as of 05:41, 8 September 2010

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Contents 1 2010/08/26(Bambi75) 2 Make Plates 2.1 member 2.2 Materials 2.3 Procedure 3 Separation of A.xylinum 3.1 member 3.2 Materials 3.3 Procedure 4 PCR 4.1 member 4.2 Materials 4.3 Procedure

2010/08/26(Bambi75)

Make Plates

member

NEX , Bambi75 and watachin

Materials

• RO water 200ml
• LB Broth 4g
• Cam(50μg/L) 10ml

Procedure

① mix materials.

② Divide ①equally and make 10 plates.

Separation of A.xylinum

member

easily and naoto

Materials

• the plate(made on August 25th).

Procedure

①extract material 2ml.

②centrifuge ① 10000rpm/5min.

PCR

member

same above

Materials

• 2×PCR buffer 25×4μl
• 2mM dNTP 10×4μl
• 10mM primer(sense)bcsA,B,C and D 2.5μl each
• 10mM primer(antisense)bcsA,B,C and D 2.5μl each
• template DNA a little
• Q water 9×4μl
• KOD FX 0.5×4μl

Procedure

①mix all materials for 4 tubes.

②elongation

• bcsA,bcsB and bcsC
  • 94℃ 2min
  • 98℃ 10sec☆
  • 55℃ 30sec
  • 68℃ 4min★
  • 68℃ 7min
  • 10℃　∞
• bcsD
  • 94℃ 2min
  • 98℃ 10sec☆
  • 55℃ 30sec
  • 68℃ 1min★
  • 68℃ 7min
  • 10℃　∞

※30cycle ☆ to ★.