The next steps are:
- Complete the PCR tests of strain genotypes and the analysis of transformation frequency data. She's still working on the PCR tests, but so far everything looks OK.
- Check the RNA concentration using the Nanodrop
- Run aliquots of the samples in a gel to check integrity of the rRNA bands (surrogate for integrity of the mRNA).
- Treat 5 µg of each sample with DNA-free. We found our stock from last year, and there's still enough to treat all our samples.
- The former RA says we can take the DNA-free-treated samples directly to the RiboXero ste; we don't need to first do a 'clean-up' step with the RNeasy Minelute kit spin-columns.
- Treat an aliquot of each sample with RiboZero. We will use only half as much RNA as recommended, and only 1/4 as much of the other reagents (in 1/4 of the recommended volume, of course). This will let us treat 24 samples with a 6-treatment RiboZero kit.
- Give the samples to the former RA in her new lab for library preparation and sequencing.