Matt Gethers/CRI, Thailand/Labwork/PCRs/Screening for Presence and Directionality of HmgA Upstream Fragment in pUC18/Gel results for 7.9.08 PCR

From OpenWetWare
Jump to: navigation, search

Gel results for 7.9.08 PCR

6.29.08

Lane 1 Lane 2 Lane 3 Lane 4 Lane 5 Lane 6 Lane 7 Lane 8 Lane 9 Lane 10 Lane 11 Lane 12 Lane 13 Lane 14
5 μl λ DNA Ladder 7.8.08 pKn003.L1 transformation colony PCR product #1 7.8.08 pKn003.L1 transformation colony PCR product #2 7.8.08 pKn003.L1 transformation colony PCR product #3 7.8.08 pKn003.L1 transformation colony PCR product #4 7.8.08 pKn003.L1 transformation colony PCR product #5 7.8.08 pKn003.L1 transformation colony PCR product #6 Space 7.8.08 pKn004.L1 transformation colony PCR product #1 7.8.08 pKn004.L1 transformation colony PCR product #2 7.8.08 pKn004.L1 transformation colony PCR product #3 7.8.08 pKn004.L1 transformation colony PCR product #4 7.8.08 pKn004.L1 transformation colony PCR product #5 pKn002.P1 negative control

100 volts, 30 minutes, 1% gel, 20 minutes staining. All lanes showed a product at ~500 bp. For pKn004, this means the ligation of the HmgR upstream fragment failed. I believe the PCR, but the fact that the vector religated to itself is surprising. Looks like my digestion is inefficient and/or my purification of digested vector from singly cut or undigested vector is shoddy. For pKn003, I'm surprised that any product showed up because the fragment to which BT188 shouldn't be present if the ligation didn't work. I have a feeling I mixed the tubes.