Team:Tokyo Metropolitan/Notebook/Fiber/2010/09/01

From 2010.igem.org

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==2010/9/1 Wednesday(Naoto)==
==2010/9/1 Wednesday(Naoto)==
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'''procedure'''
'''procedure'''
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follow to protocol3 direct PCR
follow to protocol3 direct PCR

Revision as of 04:50, 2 September 2010


Contents

2010/9/1 Wednesday(Naoto)

Experiment:digestion of bcsA,B,C,D(A.xylinus)

member

naoto

material

  • bcsA,B,C,D(from A.xylinus)
  • 10×Mbuffer
  • BSA
  • XbaI
  • SpeI

procedure

  1. add "material" to PCR tubes(show below)
  2. incubation at (37℃,7h)
composition
bcsAbcsBbcsCbcsD
solution of bcs(μl) 20202020
10×Mbuffer(μl) 2 2 2 2
BSA(μl) 2 2
XbaI(μl) 0.8 0.8
SpeI(ul) 0.8 0.8


Experiment:subculture ofA.xylinus

member

naoto

material

A.xylinus JCM7664

procedure

transfer A.xylinus JCM7664 to new culture(OWW and JCM Broth 8/25 made)


Experiment:subculture of E.coli

member

naoto

material

E.coli K12

procedure

pick up a culture of E.coli K12 and streak it to new culture(LB plate)


Experiment:direct PCR

member

naoto

material

  • E.coli K12
  • 2×PCR buffer 25μl×2
  • 2mM dNTP 10μl×2
  • 10μM primer(sense)bcsA,B 2.5μl each
  • 10μM primer(antisense)bcsA,B 2.5μl each
  • milli-Q water 9μl×2
  • KOD FX 0.5μl×2

procedure follow to protocol3 direct PCR