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From 2011.igem.org

For week three, the reporter group continued creating the lac inducible test construct. When the primers, designed during week two, arrived, the reporters began assembling the linkers and cleavage sites via PCR.

Contents 1 Monday 1.1 Lac Inducible Test: Take 2 2 Tuesday 2.1 Lac Inducible Test: Take 3

Monday

Lac Inducible Test: Take 2

Assembly Steps	Parts Involved with Assembly Step
PCR	J33204 miniprep from 5/26/11
Restriction Digest	Insert: J33204 using XbaI and PstI Vector: R0010 using SpeI and PstI
Ligation	J33204+R0010 digests
Transformation/Plating	The above ligation was transformed into Escherichia coli cells and plated on Amp resistant plates.

Tuesday

Assembly Steps	Parts Involved with Assembly Step
Colony PCR	Two colonies from plates from 5/30/11
Agarose Gel Elcectrophoresis	J33240 PCR products from 5/30/11 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 from 5/30/11.

Lac Inducible Test: Take 3

Assembly Steps	Parts Involved with Assembly Step
Restriction Digest	Insert: J33204 using XbaI and PstI Vector: R0010 using SpeI and PstI
Ligation	J33204+R0010 digests
Transformation/Plating	The above ligation was transformed into Escherichia coli cells and plated onto Amp resistant plates.