IndieBiotech:Back Door

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Here are some things I/We use at Indie Biotech. It's mostly protocols.

Contents

E.coli Transformation

By PEG/Mg. Brands verified to work: Miralax and generic "Epsom Salts".

E.coli Miniprep

Have used the Miniprep kit from NBSbio.co.uk successfully, will soon test DIY version with and without columns.

Bacillus subtilis Transformation

Still testing protocols. However, the protocol used by Spizizen in 1968 (1) is promising and appears to work well. In this method, pre-prepared spores of B.subtilis are simple resuspended in nutrient broth and transforming DNA is added. They are subjected to heat-shock at 60C and then incubated for some time before plating.

Personal experience shows that this appears to work with some efficiency (suitable for my needs but unquantified), and further work will determine whether cells require the defined minimal media rather than rich broth, or whether the heat-shock stage may be omitted if longer incubation times are used.

(1) "Transformation of biochemically deficient strains of Bacillus subtilis by deoxyribonucleate", J.Spizizen, P.N.A.S, August 4, 1968

Bacillus subtilis Miniprep

Not yet tested; presumed as above but with prior lysozyme treatment.

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