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From 2013.igem.org

MiniPreps of yesterday's liquid cultures

The plasmids were extracted from our liquid cultures, giving us DNA from...

- BBa_K592001, our green light sensor (CcaS)

- BBa_K592002, which codes for the protein (CcaR) which communicates between our green light sensor and inverter gene which eventually regulates transcription of the magenta pigment gene

- New Part 2, which has a promoter and RBS added to the FixJ coding region (BBa_K608002)

SureClean protocol

SureClean is a solution provided by bioline that allows column free nucleic acid purification. After a number of our digests we found the concentrations of resulting DNA to be unacceptably low when measured using the NanoDrop, fortunately SureClean provided us with a method of concentrating our samples.

Protocol:

1 - Add 6 ul of pink co-precipitate to your nucleic acid sample and mix well for 30s. For samples that are greater than 200ul the pink co-precipitate can be increased accordingly but it isn't ever necessary to use over 20ul.

2 - Add an equal volume of SureClean to your nucleic acid solution and mix well.

3 - Centrifuge on max speed (!4,000 x g) for 10 mins.

4 - Carefully remove supernatant by pipette.

5 - Add a volume of 70% ethanol that is equak to 2x the original sample volume and vortex for 10s.

6 - Repeat step 3.

7 - Remove supernatant and air dry to ensure all ethanol is removed.

8 - Resuspend pellet in the desired volume of TE, water or buffer.

9 - Use NanoDrop to measure concentration of resulting purified sample - hopefully it will have improved!