Alanna Greene
See Laura’s notebook entry for homogenization steps.
Realized we skipped step 3 (centrifuging again in a DNA spin column to remove DNA). We will see if we can salvage RNA, if not we will re-d0 this with different samples.
2/22/19 1:30 p.m.
Samples were removed from -80 and I began the treatment on the samples, starting at step 3 in the QIAGEN RNeasy Plus protocol. Sample 136 clogged the MiniElute column (part of homogenate did not flow through after centrifuging ). I transferred this to a new column called 136B, which I processed over to replace 136, which was completely clogged. It turns out a lot of the columns were clogged (step 6+7), so at step 7 I increased the speed to 16G in the centrifuge. Once the protocol was completed, I stored the samples in the fridge.
3/1/9- RNA QUANTIFYING
- Autoclaved dishes and got more liquid nitrogen
- Ran samples through Qubit 30 following protocol:
- Wiped down tubes
- Calibrated standards first (blank and full)
- For samples: 1 microliter in sample in 199 microliter in solution
- Successfully quantified!
| Sample # | Qubit Reading |
| S1 | Too low out of range |
| S2 | 9.92 ng/mL |
| 136 | 116 |
| 112 | 192 |
| 100 | Too high |
| 124 | Too high |
| 115 | 98.2 |
| 106 | 110 |
| 126 | Too high |
| 140 | 102 |
Roberts Lab 