User:SBurke
Problems with PCR using VR/VF2
Robotic Assembly
- Assembly was the same as the one on 6/7/2007 except it used the old construction plasmid (prepped by ML on 3.13)
- Tested transformation of 1ul of pSB1AK3 (LIGATED) in 50ul cells, plated all out on AK plates
- Transform 1ul into 50ul cells (Top10-0008), do transformation on thermal module. Plate all 251ul on correct antibiotic plates.
- Began counting colonies at 12:45PM
| Construct color |
Assembly construct | Number of colonies 1ul ~22hrs |
|
|---|---|---|---|
| RFP | A. 1AC3.J04500.I13501 | 167/184 | |
| GFP | B. 1AC3.R0010.I13504 | 80/91 | |
| RFP | C. 1AT3.J04500.I13501 | 41/61 | |
| GFP | D. 1AT3.R0010.I13504 | 24/40 | |
| RFP | E. 1AK3.J04500.I13501 | 18/811 | |
| GFP | F. 1AK3.R0010.I13504 | 0 | |
| X-S control | 69 (250ul plated) | ||
| pUC control (in lig prod as usual) | 0 (250ul plated) | ||
| RFP J04450 | 734/734 (plated 50ul; 250ul plating yields 3670) | ||
| pSB1AK3 (ligated) | 41 (250ul plated) |
| Assembly Plate | Resulting construct | Number of colonies 1ul ~22hrs |
|---|---|---|
| 2A | BBa_P200714 | 1 |
| 2B | BBa_P20060 | 2 |
| 2C | BBa_Q200514 | 7 |
| 2D | BBa_B3210 | 41 |
| 2E | BBa_B3210 | 20 |
| 2F | BBa_B3209 | 9 |
| 2G | BBa_B3203 | 17 |
| 2H | BBa_B3202 | 1 |