Team:SVCE CHENNAI/Restriction

Lactoshield - SVCE_CHENNAI Lactoshield - SVCE_CHENNAI

Lactoshield - SVCE_CHENNAI Lactoshield - SVCE_CHENNAI

Restriction


Requirements for each sample:

  •   Ice and bucket/container
  •   1) 8-tube strip, or (3) 0.6ml thin-walled tubes
  •   BioBrick Part in BioBrick plasmid (Purified DNA, > 16ng/ul)
  •  dH2O
  •   NEB Buffer 2
  •  BSA
  •   Restriction Enzymes: EcoRI, SpeI, XbaI, PstI
  •  Thermal cycler

Procedure:

  •   Add 250ng of DNA to be digested, and adjust with dH20 for a total volume of 16ul.
  •   Add 2.5ul of NEBuffer 2.
  •   Add 0.5ul of BSA.
  •  Add 0.5ul of EcoRI.
  •   Add 0.5ul of PstI.
  •  There should be a total volume of 20ul. Mix well and spin down briefly.
  •  Incubate the restriction digest at 37C for 30min, and then 80C for 20min to heat kill the enzymes.
  •  Run a portion of the digest on a gel (8ul, 100ng), to check that both plasmid backbone and part length are accurate.
Lactoshield - SVCE_CHENNAI Lactoshield - SVCE_CHENNAI
Lactoshield - SVCE_CHENNAI

Sponsors


igemsvce@gmail.com

Sri Venkateswara College of Engineering
Tamil Nadu, India

    

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