Team:KAIT Japan/Protocol

From 2013.igem.org

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='''Protocol'''=
='''Protocol'''=
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=='''Miniprep'''==
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#Add culture medium 1mL in a microtube
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#Centrifuge(1min,4°C,10,000rpm)
 +
#Remove the supernatant to new microtube
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#Repeat 1-3
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#Add SolI 100μL and Vortex
 +
#Add SolII 200μL and invert
 +
#ice-cold 3min
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#Add SolIII 150μL and invert
 +
#ice-cold 3min
 +
#Centrifuge(10min,4°C,10,000rpm)
 +
#Add the supernatant to new microtube
 +
#Add RNase 0.8μL
 +
#Incubation(1h,37°C)
 +
#Add phenol:chloroform 200μL
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#Vortex
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#Centrifuge(5min,4°C,10,000rpm)
 +
#Add the supernatant to new microtube
 +
#Add chloroform 200μL
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#Tapping
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#Centrifuge(1min,4°C,10,000rpm)
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#Add the supernatant(150μL) to new microtube
 +
#Add 3M-acetic acid 15μL
 +
#Add 100%Et 400μL and invert
 +
#Centrifuge(20min,4°C,10,000rpm)
 +
#Remove the supernatant to new microtube
 +
#Add 70%Et 400 μL
 +
#Tapping
 +
#Centrifuge(20min,4°C,10,000rpm)
 +
#Remove the supernatant to new microtube
 +
#Dry
 +
#Add TE buffer 50μL
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#Storage

Revision as of 02:52, 19 August 2013

KAIT Japan 2013RogoTop.png

KAIT Japan2013 Kanako.png KAIT Japan 2013 iGEMRogo.png

KAIT Japan home.png Home
KAIT Japan team.png Team
KAIT Japan project.png Project
KAIT Japan parts.png Parts

KAIT_Japan_protocol.png Protocol

KAIT Japan notebook.png Notebook
KAIT Japan results.png Results
KAIT Japan safety.png Safety
KAIT Japan human practice.png Human Practice


Protocol

Miniprep

  1. Add culture medium 1mL in a microtube
  2. Centrifuge(1min,4°C,10,000rpm)
  3. Remove the supernatant to new microtube
  4. Repeat 1-3
  5. Add SolI 100μL and Vortex
  6. Add SolII 200μL and invert
  7. ice-cold 3min
  8. Add SolIII 150μL and invert
  9. ice-cold 3min
  10. Centrifuge(10min,4°C,10,000rpm)
  11. Add the supernatant to new microtube
  12. Add RNase 0.8μL
  13. Incubation(1h,37°C)
  14. Add phenol:chloroform 200μL
  15. Vortex
  16. Centrifuge(5min,4°C,10,000rpm)
  17. Add the supernatant to new microtube
  18. Add chloroform 200μL
  19. Tapping
  20. Centrifuge(1min,4°C,10,000rpm)
  21. Add the supernatant(150μL) to new microtube
  22. Add 3M-acetic acid 15μL
  23. Add 100%Et 400μL and invert
  24. Centrifuge(20min,4°C,10,000rpm)
  25. Remove the supernatant to new microtube
  26. Add 70%Et 400 μL
  27. Tapping
  28. Centrifuge(20min,4°C,10,000rpm)
  29. Remove the supernatant to new microtube
  30. Dry
  31. Add TE buffer 50μL
  32. Storage





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