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mm/dd/yyyy
Ran PCR products from yesterday on a gel. 5ul of reactions 2-9, all 30 of 1 so I can gel extract because there should be two bands and I only want one.
DpnI digest PCR reactions 2-7 and 9 with positive and negative controls (didn't run reaction 8 because I checked the gel early and saw that there was no DNA there).
Controls
Positive control was a miniprepped plasmid at 100ng/ul concentration.
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Volume (ul)
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'
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| Thing |
Positive ctrl |
Negative ctrl
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| H2O |
12 |
12
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| Green digest buffer |
2 |
2
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| pTet mCh |
5 |
5
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| DpnI |
1 |
0
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| Thing
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Volume (ul)
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| Digest buffer |
4
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| PCR rxn |
25
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| H2O |
1
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| DpnI |
1
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Incubated at 37°C for 55 minutes, sat on the bench for 3 minutes, heat inactivated at 80 for 5 minutes.
Cleaned using zymo DNA clean and concentration-5 kit (columns from gel extraction kit). Added 150ul of DNA binding buffer to DpnI reactions.
Gel extracted PCR reaction 1. Saw a smaller band above, think I got the correct band.
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