BglII digest followed by PCR and agarose gel extraction #
Digesting vector bound VR fragments with BglII then amplifying by PCR. Digest conditions are listed below.
- 1 ul (50 ng vector)
- 1 ul 10x 3.1 buffer
- 0.2 ul BglII
- 7.8 ul npH20
After 2 hours of digestion the samples were then amplified using a PCR master mix made up of lab Taq, dNTP, OneTaq PCR buffer and VR inert primers.
Agarose gel extraction #
After PCR loaded 2 ul of each sample into top rows of wells for visualization. Loaded remaining samples into corresponding bottom row well.
Lane layout #
After PCR completed; 11th lane is a control with no insert added.
| Lane | Sample |
|---|---|
| 1 | 8 |
| 2 | 12 |
| 3 | 15 |
| 4 | 16 |
| 5 | 17 |
| 6 | 21 |
| 7 | 23 |
| 8 | 25 |
| 9 | 27 |
| 10 | 31 |
| 11 | C |
Control lane shows contamination bottom fuzzy lanes that are present in the gel. That is what we call not good. Need to run negative control reactions to figure out what is going on here.