Team:NTNU Trondheim/Protocols

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NTNU IS B.A.C.K.
Bacterial Anti-Cancer-Kamikaze

Protocols

This is a list of recipes and protocols used by the team.

Contents

Transformation

Official iGEM transformation protocol: http://partsregistry.org/Help:Protocols/Transformation

We use this protocol with the following modifications:

The cells used are Subcloning Efficiency™ DH5α™ Competent Cells from Life Technologies/Invitrogen

Inoculation after transformation

Using a sterile toothpick, pick/scratch a single colony from the transformants. Drop the toothpick into a plastic tube with 3 mL sterile liquid medium with the appropriate antibiotic(s). Close the tube but leave the cap slightly open to allow oxygen to enter, and incuate at 37 C with shaking.

DNA Isolation

We use the Promega Wizard Plus SV Minipreps DNA Purification System A1460 with the sentrifugation version of the protocol supplied by the vendor.

DNA Concentration measurements

Concentrations of DNA after isolation was measured with the NanoDrop ND-1000 Spectrophotometer.

Restriction digest

We are using the single reaction protocol from partsregistry.org : [1]

Ligation

Ligation protocol from partsregistry.org [2]:

In stead of incubating as described above, the samples are kept at 37 degrees celsius in a water bath for 60 minutes.

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