PRODUCT: PRECIPITATION CARRIER

Cat. No: PC 173
Storage: Store at 4C

PRODUCT DESCRIPTION

Precipitation Carrier is a molecular biology grade solution of acryl polymer designed for use in the isolation of small amounts of RNA (< 10 g). It can be used with both RNAzol® RT and TRI Reagent®. In the RNAzol RT procedure, Precipitation Carrier facilitates the recovery of both large (> 200 bases) and small (< 200 bases) RNA in separate fractions. This carrier does not affect the activity of restrictases, reverse transcriptase, Taq polymerase, ligase or other enzymes used for nucleic acid analysis. Precipitation Carrier can be used for at least one year when stored at room temperature or 4 C. The shelf life can be extended beyond one year by storage at -20 C.

APPLICATION

1. RNAzol® RT - Isolation of large RNA (> 200 bases). Add 1 - 3 µl of Precipitation Carrier when using less than 1.0 ml of reagent, or 3 - 5 µl of carrier for more than 1.0 ml of RNAzol RT. Perform sample homogenization or lysis, add Precipitation Carrier and isolate the RNA as described in the RNAzol RT protocol. Alternatively, add Precipitation Carrier to the water - supernatant obtained after the removal of DNA and proteins (Step 3, RNAzol RT protocol). Add 0.4 volumes of 75% ethanol, briefly mix and store at room temperature for 5 - 10 minutes. Sediment RNA precipitate at 12,000 g for 8 minutes at 4 - 25 C. Dissolve the precipitate by repetitive pipetting in DEPC - treated water or other solubilizing medium. Please refer to the RNAzol RT protocol for details.

2. RNAzol® RT - Isolation of small RNA (< 200 bases). Homogenize or lyse the sample in RNAzol RT and obtain the water - supernatant after the removal of DNA and proteins. Perform large RNA precipitation by adding 0.4 volumes of 75% ethanol, mix and sediment the large RNA. Preserve the water - ethanol - supernatant for small RNA isolation. Add 1 - 3 µl of Precipitation Carrier to this supernatant and mix. Add 0.8 volumes of isopropanol, mix and store for 30 minutes at 4 C. Sediment small RNA at 12,000 g for 15 minutes at 4 - 25 C. Dissolve the precipitate by repetitive pipetting in DEPC - treated water or other solubilizing medium. Please refer to the RNAzol RT protocol for details.

3. TRI Reagent® Isolation of total RNA. Add 1 - 3 µl of Precipitation Carrier when using less than 1.0 ml of reagent, or 3 - 5 µl of carrier for more than 1.0 ml of TRI Reagent. Perform sample homogenization or lysis and add Precipitation Carrier to the homogenate. Isolate total RNA as described in the TRI Reagent protocol. Alternatively, add Precipitation Carrier to the aqueous phase obtained after phase separation and mix. Add isopropanol and proceed with RNA isolation as described in the TRI Reagent protocol. Please refer to the TRI Reagent protocol for details.

NOTES. Precipitation Carrier contributes to the optical density of RNA preparations. To normalize for this effect, process a blank sample containing only the reagent and Precipitation Carrier. Solubilize the final pellet of the carrier in the same volume of solubilization solution used for the test sample pellets. Measure the optical density at 260 and 280 nm for all samples and the blank, and subtract the values of the blank from values obtained for each RNA sample.

MRC GUARANTEE

RNase activity - none detected.