Team:Newcastle/Labwork/21 September 2009

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Formal Lab Session - 21st September 2009

Stochastic switch team

Today we digested the sac minipreps with EcoRI and XbaI. It seems clear that there was something wrong with the minipreps (such as poor procedure) as there seemed to be little or no DNA in the sample.

Did a gel fragment clean up on the sspb fragment cut from the gel, this was put in the 'fragments' box in the freezer ready for later.

50ul of ara+pSB1AT3 plasmid was cut with SpeI and PstI in order to insert the sspb fragment after the operator. Rather than doing a gel fragment prep a PCR clean up (kit) was done on the digest to save time.

We ligated the ara + sspb fragments using Chris' method:

  • 2ul Ligase buffer
  • 2.5ul Vector DNA
  • 14.5ul insert DNA
  • 1ul ligase

= 20ul

This was left in the fridge overnight to improve the ligation conditions.




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