E.Z.N.A.® Total RNA Kit I

Can samples be stored in TRK Lysis Buffer prior to extraction?

Homogenized tissue/cell lysates in TRK Lysis Buffer can be stored at -80 °C for at least 6 months. To proceed with the frozen lysates, thaw the sample at 37 °C until they are completely thawed, and all salts in the lysis buffer are dissolved. Do not extend the treatment in 37 °C because it can cause chemical degradation of RNA.

I have been getting low 260/230 ratios with my samples. Is there a way to improve this?

Low A260/A230 ratio alludes to salt carryover (guanidine salt is not completely washed off during wash steps). Typically increasing the wash steps improves this ratio – so instead of 2 RNA Wash Buffer II wash steps, do 3.

Is there a way to isolate miRNA using the E.Z.N.A. Total RNA Kit I?

To bind RNA <200 nt, they would need to modify the binding step by adding 1 volume of 100% ethanol (instead of 70% ethanol in Step 5, Page 14, and Step 5, Page 19), and rest of the steps remain as-is. This modification helps bind miRNA along with large RNA and will get co-extracted in the same fraction.

Can I use the E.Z.N.A. Total RNA Kit I for tissues or cells stored in RNAlater™?

The RNAlater will interfere with our buffer chemistry resulting in low yields, so tissues must be washed prior to starting the extraction.

1. Spin down the tissue at 500 x g for 5 minutes.
2. Aspirate out the supernatant.
3. Spin down the sample again (500 x g for 5 minutes), then aspirate out the supernatant.
4. Use the pelleted cells or tissue for extraction following the E.Z.N.A. Total RNA Kit I Protocol – Culture Cells on page 11 or E.Z.N.A. Total RNA Kit I Protocol – Animal Tissue on page 17.