Team:UNICAMP-EMSE Brazil/protocols/Digestion

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==Digestion==
==Digestion==
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(For each 1000 ng of DNA use 1 U of enzyme in 20uL reaction)
+
 
-
Use 3000 ng BioBrick vector  
+
(For each 1000 ng of DNA use 1 U of enzyme in 20uL reaction)
-
6 µL 10x buffer
+
#Use 3000 ng BioBrick vector  
-
0.5 µL enzyme 1 (20 units/µL)
+
#6 µL 10x buffer
-
0.5 µL enzyme 2 (20 units/µL)
+
#0.5 µL enzyme 1 (20 units/µL)
-
distilled water to 60 µL total volume
+
#0.5 µL enzyme 2 (20 units/µL)
-
+
#distilled water to 60 µL total volume
 +
 
Put in 0,2 mL eppendorf tubes and let 2 h at 37°C.
Put in 0,2 mL eppendorf tubes and let 2 h at 37°C.
Make an electrophoresys (use the hole reaction volume) and cut the band with your gene of interest.
Make an electrophoresys (use the hole reaction volume) and cut the band with your gene of interest.

Revision as of 20:46, 22 August 2011

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Digestion

(For each 1000 ng of DNA use 1 U of enzyme in 20uL 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 electrophoresys (use the hole reaction volume) and cut the band with your gene of interest.
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