26/07/13

From 2013.igem.org

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Revision as of 16:51, 26 July 2013

purification of the limonene biobrick - BBa_24

Plasmid Volume

-1.1 - 40ul -1.2 - 40ul -1.3 - 46ul -1.4 - 39ul -1.5 - 41ul -1.2 - 32ul

measured DNA concentration using nanodrop
DNA concentration (ng)

-1.1 - 131.8 -1.2 - 77.1 -1.3 - 56.8 -1.4 - 109 -1.5 - 129.1 -1.2 - 93.3

Restriction enzyme digest of the DNA samples

-enzymes: XbaI, PstI -master mix: Buffer - 14ul

            H2O - 88.2ul
            XbaI - 1.4ul
            PstI - 1.4ul
    Total volume - 105ul

Each sample contained 5ul of DNA and 15ul of the master mix. The total amount of DNA in each sample was 200ng To calculate it we divided 200ng by the concentration of each sample The rest of the volume was made up with water to give a total of 5ul. The samples were them incubated in a water bath at 37C by an hour

Agorose Gel preparation

-20ml of 5xTBE -80ml of water -0.8g of agarose

Boil for 3 minutes, 1 minute at a time. Let it cool and add 5um of ethidium bromide. Pour and let set for 30 mins.

Adding dye and and marker

Used orange g loading dye. -2um to each sample

Used 1kb ladder, 5um per 1 marker well.

Run the gel

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 purification of the limonene biobrick - BBa_24
 *Plasmid Volume