Team:Newcastle/Labwork/21 August 2009

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Contents 1 Stochastic Switch team 1.1 Summary 1.2 Miniprep of C0178 cultures 1.2.1 N.B: During our minipreps we have always kept solution III on the bench, however Chris seeing this advised us to always keep solution III in the fridge for the best results. As well as this after the addition of solution III the tubes must be shaken well in order for the proteins to precipitate properly. Remember that Phil's protocols may not always be explicit and although care should be taken in following them, there may be important points missed out that can effect results.

Stochastic Switch team

Summary

Today the stochastic team split into two, to fully utilise the time in the lab. Jess carried out minipreps for the BBa_C0178 in E.coli DH5alpha cultures that were set up yesterday. Goksel carried oud restriction digests of the BioBricks R0062; R0079; C0161; C0179; J44000 which were miniprepped yesterday.

Miniprep of C0178 cultures

As it happens only 2 of the 3 cultures grew from yesterdays overnights so these were spun down (1ml of each). It was decided that these cultures would be treated separately until after being run on a gel so their identities could be confirmed.

After spinning the cultures down after the addition of solution III, it was observed that there was not much of a protein pellet to be seen which should precipitate after solution III is added. The tubes were shaken well and put in to centrifuge for a further 10 miutes and a larger pellet was seen.

N.B: During our minipreps we have always kept solution III on the bench, however Chris seeing this advised us to always keep solution III in the fridge for the best results. As well as this after the addition of solution III the tubes must be shaken well in order for the proteins to precipitate properly. Remember that Phil's protocols may not always be explicit and although care should be taken in following them, there may be important points missed out that can effect results.

The supernatant was taken and Isopropanol added, the solution was spun, ethanol added, aspirated and speed vac'd according to the protocol. The DNA pellet was resuspended in 50ul water, labelled, and frozen in the -20 freezer.