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|