Matt Gethers/CRI, Thailand/Labwork/Gels/Week of 6.29.08

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Contents

6.29.08

Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 Lane 7 Lane 8 Lane 9
5 μl λ DNA Ladder5 μl 2736/2737 Positive Control (genomic DNA template, Taq) 50oC5 μl 2736/2737 Positive Control (genomic DNA template, Taq)5 μl 2736/2737 Positive Control (genomic DNA template, Taq)5 μl 2736/2737 Positive Control (genomic DNA template, Taq)5 μl 2736/2737 Positive Control (genomic DNA template, Taq)5 μl 2736/2737 Positive Control (genomic DNA template, Taq)5 μl 2736/2737 Positive Control (genomic DNA template, Taq)5 μl 2736/2737 Positive Control (genomic DNA template, Taq) 65o

100 volts, 30 minutes, 1% gel, 20 minutes staining. This was a gradient PCR to determine the optimum annealing temperature. All lanes showed product (50 to 65 degrees).

6.30.08

Gel #1

Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 Lane 7 Lane 8 Lane 9
5 μl λ DNA Ladder5 μl 2736/2737 6.27.08 pKn001.1 transformation colony PCR Product #1 (Taq)5 μl 2736/2737 6.27.08 pKn001.1 transformation colony PCR Product #2 (Taq)5 μl 2736/2737 6.27.08 pKn001.1 transformation colony PCR Product #3 (Taq)5 μl 2736/2737 6.27.08 pKn001.1 transformation colony PCR Product #4 (Taq)5 μl 2736/2737 6.27.08 pKn001.1 transformation colony PCR Product #5 (Taq)5 μl 2736/2737 6.27.08 pKn001.1 transformation colony PCR Product #6 (Taq)5 μl 2736/2737 6.27.08 pKn001.1 transformation colony PCR Product #7 (Taq)5 μl 2736/2737 Positive Control (genomic DNA template, Taq)

100 Volts, 30 minutes, 1% gel, stained for 20 minutes. The positive control worked beautifully and it looks like colony 3 turned up a product at the correct length.


Gel #2

Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 Lane 7 Lane 8 Lane 9
5 μl λ DNA Ladder5 μl M13_for/BT2696 6.27.08 pKn002.2 transformation colony PCR Product #1 (Taq)5 μl M13_for/BT2696 6.27.08 pKn002.2 transformation colony PCR Product #2 (Taq)5 μl M13_for/BT2696 6.27.08 pKn002.2 transformation colony PCR Product #3 (Taq)5 μl M13_for/BT2696 6.27.08 pKn002.2 transformation colony PCR Product #4 (Taq)5 μl M13_for/BT2696 6.27.08 pKn002.2 transformation colony PCR Product #5 (Taq)5 μl M13_for/BT2696 6.27.08 pKn002.2 transformation colony PCR Product #6 (Taq)5 μl 2736/2737 6.27.08 pKn002.2 transformation colony PCR Product #7 (Taq)5 μl M13_for/BT2696 Positive Control (genomic DNA template, Taq)

100 Volts, 30 minutes, 1% gel, stained for 20 minutes. Nothing showed up at all, including the positive control.

7.2.08

Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 Lane 7 Lane 8 Lane 9
5 μl λ DNA Ladder5 μl M13_rev/BT1033 6.27.08 pKn002.2 transformation colony PCR Product #1 (Taq)5 μl M13_rev/BT1033 6.27.08 pKn002.2 transformation colony PCR Product #2 (Taq)5 μl M13_rev/BT1033 6.27.08 pKn002.2 transformation colony PCR Product #3 (Taq)5 μl M13_rev/BT1033 6.27.08 pKn002.2 transformation colony PCR Product #4 (Taq)5 μl M13_rev/BT1033 6.27.08 pKn002.2 transformation colony PCR Product #5 (Taq)5 μl M13_rev/BT1033 6.27.08 pKn002.2 transformation colony PCR Product #6 (Taq)5 μl M13_rev/BT1033 6.27.08 pKn002.2 transformation colony PCR Product #7 (Taq)5 μl M13_rev/BT1032 Positive Control (genomic DNA template, Taq)

100 Volts, 30 minutes, 1% gel, stained for 20 minutes. Every lane had some product, but there were usually multiple bands (some at ~600, then 1500, and one higher, I think). Of all the colony PCR lanes, 1 and 2 had the brightest bands at ~600.

7.4.08

Lane 1 Lane 2
5 μl λ marker20 μl pKn002.P1 HindIII/SphI digest

100 volts, 30 minutes, 1% agarose. Stained in EtBr for 15 minutes. Excised band at ~4.2 Kb.

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