Dewaxing Solution

The CELLDATA RNAstorm™ 2.0 FFPE RNA Extraction Kit (Cat. No. CD506) and CELLDATA DNAstorm™ 2.0 FFPE DNA Extraction Kit (Cat. No. CD507) contain a Dewaxing Solution that was reformulated for IP considerations. These kits are direct replacements for the discontinued CELLDATA RNAstorm™ or DNAstorm™ FFPE Extraction Kits (CD501, CD502). The kits continue to utilize Biotium’s proprietary CAT5™ catalytic technology for efficient reversal of formaldehyde crosslinks.

The maximum capacity of the spin columns in the kit is similar to a standard miniprep column, about 20 ug of DNA. However, the expected yield from FFPE extraction is much lower, it is very rare to get more than 1-2 ug of DNA per prep.

Yes, the RNAStorm™ and DNAStorm™ FFPE kits may be used sequentially. The steps below will allow the protocol to be adapted to extract both RNA and DNA from one sample. You can also download this information in an app note.

Begin by extracting the sample according to the RNAstorm™ kit protocol with the following modifications:

1.  Perform step 3 (normally a 2 hour incubation) for only 30 minutes at 72?C. See note below regarding possible optimization of this step.
2.  Perform steps 4 and 5 of the RNAstorm™ protocol as directed, but do not discard the pellet (which contains the DNA) in step 5.
3.  Transfer the supernatant (which contains the RNA) to a new tube as instructed in step 6.
4.  Continue to incubate the supernatant for another 1.5 hours at 72?C (2 hours total including the initial 30 minutes), then proceed with step 7 of the RNAstorm™ protocol (add Binding Buffer) and all remaining steps as instructed.
5.  Use the pellet from step 2, which contains DNA, as input for step A5 (or B8, depending on deparaffinization choice) of the DNAstorm™ kit manual.
6.  Continue with step A5 (or B8) of the DNAstorm™ protocol by adding 200 µL of CAT5™ Buffer to the pellet, then continue as instructed by the DNAstorm™ protocol.

Note: the initial incubation period can be adjusted depending on relative DNA and RNA yields. If the RNA yield is high but the DNA yield is low, reduce the incubation time in step 3 (no less than 15 mins). If the DNA yield is good but the RNA yield is low, increase the incubation time in step 3 (no more than 2 hours).