Team:UNICAMP-EMSE Brazil/protocols/Digestion

From 2011.igem.org

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[[Image:UNICAMP-EMSE_Brazil_E_Coli.png|270px|right]]
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[[Image:UNICAMP-EMSE_Brazil_Restriction.png|270px|right]]
==<font size="5" color=#086A87>Digestion:==
==<font size="5" color=#086A87>Digestion:==
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(For each 1000 ng of DNA use 1 U of enzyme in 20uL reaction)
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(For each 1000 ng of DNA use 1 U of enzyme in 20µL reaction)
#Use 3000 ng BioBrick vector  
#Use 3000 ng BioBrick vector  
#6 µL 10x buffer
#6 µL 10x buffer
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#distilled water to 60 µL total volume
#distilled water to 60 µL total volume
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Put in 0,2 mL eppendorf tubes and let 2 h at 37°C.
 
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Make an electrophoresys (use the hole reaction volume) and cut the band with your gene of interest.
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*'''Put in 0,2 mL eppendorf tubes and let 2 h at 37°C.'''
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*'''Make an electrophoresis (use the hole reaction volume) and cut the band with your gene of interest.'''

Latest revision as of 00:58, 23 August 2011

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UNICAMP-EMSE Brazil Restriction.png

Digestion:

(For each 1000 ng of DNA use 1 U of enzyme in 20µL reaction)

  1. Use 3000 ng BioBrick vector
  2. 6 µL 10x buffer
  3. 0.5 µL enzyme 1 (20 units/µL)
  4. 0.5 µL enzyme 2 (20 units/µL)
  5. distilled water to 60 µL total volume


  • Put in 0,2 mL eppendorf tubes and let 2 h at 37°C.
  • Make an electrophoresis (use the hole reaction volume) and cut the band with your gene of interest.






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