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From 2011.igem.org

Contents 1 Monday 1.1 Reporters 1.1.1 PCR 1.1.2 Restriction Digest 1.1.3 Ligation 1.1.4 Transformation/Plating 2 Tuesday 2.1 Reporters 2.1.1 Colony PCR 2.1.2 Agarose Gel Electrophoresis 2.1.3 Restriction Digest 2.1.4 Ligation 2.1.5 Transformation/Plating 3 Wednesday

Monday

Reporters

PCR

     J33204 miniprep from 5/26/11

Restriction Digest

• Insert with XbaI and PstI

     J33204

• Vector with SpeI and PstI

     R0010

Ligation

     J33204+R0010 digests

Transformation/Plating

     The above ligation was transformed into Escherichia coli cells and plated on Amp resistant plates.

Tuesday

Reporters

Colony PCR

     Two colonies from plates from 5/30/11

Agarose Gel Electrophoresis

1. J33240 PCR products
2. Colony PCR products from above

• Results: The gel showed that the insert (J33240) PCR worked, but the assembly did not work. We came to this conclusion because the constructs on the gel were less than 500 base pairs and our desired construct should have been about 1100 base pairs. Thus, we needed to do assembly for a third time. We started this third assembly with the R0100 miniprep from 5/26/11 and the J33204 PCR products.

Restriction Digest

• Insert with XbaI and PstI

     J33204

• Vector with SpeI and PstI

     R0010

Ligation

     J33204+R0010 digests

Transformation/Plating

     The above ligation was transformed into Escherichia coli cells and plated onto Amp resistant plates.

Wednesday

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