IGEM:UC Berkeley/2006/Digestion

Digestion of a Mini-prep DNA (35 ul scale)

1. 13 uL water
2. 3.5 uL buff(3)
3. 3.5 uL BSA
4. 1 uL Enzyme 1
5. 1 uL Enzyme 2
6. 15 uL DNA

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  35 

INCUBATE AT 37 degrees for 2 hours!

Digestion of a Mini-prep DNA (20 uL scale)

1. 10 uL DNA
2. 7 uL water
3. 2 uL 10x buffer
4. 0.5 uL Enzyme 1
5. 0.5 uL Enzyme 2

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  20 

Scaled up digestion of mini-prep

1. 14uL H2O
2. 4uL NEB Buffer 2
3. 20uL DNA
4. 1uL restriction enzyme
5. 1uL restriction enzyme

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   40 

Digestion of PCR Product

1. Elute 108uL of water
2. 12 uL of 10X NEB Buffer 2
3. 1 uL Enzyme 1
4. 1 uL Enzyme 2

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  120 uL