Team:UNICAMP-EMSE Brazil/protocols/Digestion
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{{Template:UNICAMP-EMSE Brazil_Main_Menu}} | {{Template:UNICAMP-EMSE Brazil_Main_Menu}} | ||
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+ | <font size="4" color="red">Back to [https://2011.igem.org/Team:UNICAMP-EMSE_Brazil/protocols Protocols ↑]</font> | ||
+ | </div> | ||
- | ==Digestion== | + | [[Image:UNICAMP-EMSE_Brazil_Restriction.png|270px|right]] |
+ | ==<font size="5" color=#086A87>Digestion:== | ||
+ | </font> | ||
- | (For each 1000 ng of DNA use 1 U of enzyme in | + | (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.''' | |
+ | *'''Make an electrophoresis (use the hole reaction volume) and cut the band with your gene of interest.''' | ||
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Digestion:
(For each 1000 ng of DNA use 1 U of enzyme in 20µL reaction)
- Use 3000 ng BioBrick vector
- 6 µL 10x buffer
- 0.5 µL enzyme 1 (20 units/µL)
- 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.
- Make an electrophoresis (use the hole reaction volume) and cut the band with your gene of interest.