Mini prep VR insert cultures #
Came in today to mini prep the cultures I started yesterday. I followed the lab mini prep protocol. One thing to note for future preps is that the protocol is expecting you to be working with 2 ml tubes the whole time, if you start with more than 2ml of culture (I used 8ml to get more DNA) cultures transfer cultures from glass tubes to 15 ml centrifuge tubes. Spin down cells in those cells and re-suspend, then transfer to 2ml tubes.
Mini prep results #
| Sample | Colony | Yield |
|---|---|---|
| VR-1 | 1 | 2316 |
| VR-1 | 2 | 527 |
| VR-1 | 3 | 2302 |
| VR-1 | 4 | NA |
| VR-1 | 5 | 2115 |
| VR-1 | 6 | NA |
| VR-1 | 7 | NA |
| VR-10 | 1 | 2371 |
| VR-11 | 2 | NA |
| VR-11 | 3 | NA |
| VR-11 | 4 | NA |
| VR-11 | 5 | NA |
| VR-14 | 1 | NA |
| VR-14 | 2 | NA |
| VR-14 | 3 | 2023 |
| VR-14 | 6 | NA |
| VR-14 | 7 | NA |
| VR-2 | 1 | NA |
| VR-2 | 2 | NA |
| VR-2 | 3 | NA |
| VR-2 | 4 | 1993 |
| VR-2 | 5 | NA |
| VR-2 | 6 | NA |
| VR-2 | 7 | 1859 |
| VR-4 | 1 | NA |
| VR-4 | 2 | 2310 |
| VR-4 | 2 | NA |
| VR-4 | 2 | NA |
| VR-4 | 5 | NA |
| VR-5 | 1 | NA |
| VR-5 | 2 | NA |
Samples were suspended in 100 ul of TE. There was obviously a sample mixup when I transfered from the 15ul tubes to 2ml because there are 4 VR-4 colony 2 samples. Will require sequencing to determine exactly what is in these. Like the last plasmid prep it seems that many of the samples are contaminated with RNA. Will need to run all samples on gel to confirm and then purify with LiCl protocol that is described as part of the lab midi-prep protocol. The table is also available as a csv file at this link.
Sample storage #
I started a new box for mini prep samples called VR insert midi preps and
add these samples to that box which is stored on my shelf in the the kitchen
freezer.
One Monday will do LiCl clean up of samples and then run on gel. Also prepare 2 colony extracts of each insert and send for sequencing. Plan Gibson assembly (spreadsheet, prepare reagents and label tubes) for Tuesday.