RNA Isolation & Quantification - C.bairdi RNA from Sample 6129_403_26

Since I was isolating gDNA from C.bairdi 6129_403_26 hemolymph, I figured I might as well co-isolate RNA since I was using the Quick DNA/RNA Microprep Plus Kit (ZymoResearch).

Followed the manufacturer’s protocol with the following notes/changes:

• Used 480uL of sample, combined with 1:1 H₂O (480uL), with 4:1 Lysis Buffer (3840uL) and mixed in 15mL conical

• Flow-through was retained in a 15mL conical for RNA isolation

• Column began to clog during loading of the initial flow-through + EtOH mixture

• Eluted with 15uL

Quantified on the Roberts Lab Qubit 3.0 using the high-sensitivity RNA Assay (Invitrogen) and 1uL of sample.

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RESULTS

Qubit results (Google Sheet):

• 20200210_qubit_crab_RNA

[RNA] = 7.86ng/uL in 15uL

Yield = ~118ng

Yield is very, very low, considering the amount of starting material. Yield is likely due to clogging of column, due to overloading.

Sample was stored at -80^oC in:

Rack 2, 3, 5 in Shellfish Box #8