Restricted

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Restricting DNA

We have conducted restrictions in varying concentrations and total volumes; however they all follow the same basic procedure. Below are two of the different restrictions we carried out.

Dilute

  • 46μl MillQ H2O
  • 10μl 10 x buffer
  • 40μl plasmid sample
  • 2μl enzyme 1
  • 2μl enzyme 2

Total volume = 100μl

Concentrated

  • 10μl MilliQ H2O
  • 3μl 10 X buffer
  • 10μl plasmid sample
  • 1μl enzyme 1
  • 1μl enzyme 2

Total volume = 30μl

Incubate solutions for 90 minutes in a 37°C water bath.

If DNA is to be run on a gel and a band cut out, this can be done without purifying the DNA. For all other downstream applications or for storage, solution must be purified to remove the enzymes.

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