Trizin for RNA Isolation

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 Description

Trizin enables the isolation of RNA by a single step color guided biphasic extraction method from biological samples of different sources (e.g., human, other animals, plants, yeast, bacterial and virus origin, etc.). The isolated RNA is substantially free of genomic DNA and hence ready for Reverse Transcriptase PCR reaction. The bright purple color of the reagent makes the phase separation (purple colored organic phase and colorless aqueous phase) clearly visible. Color guided separation not only minimizes the chance of getting contamination during the collection of aqueous phase but also offers a substantial yield in RNA isolation.

 Use in Downstream Application

  Isolated RNA can be used in RT PCR, Northern Blot analysis, Dot Blot hybridization, poly(A)+ selection, in vitro translation, RNase protection assay, and molecular cloning. Isolated DNA can be used in PCR, Restriction Enzyme digestion, and Southern Blots Isolated protein can be used for Western Blots, recovery of some enzymatic activity, and some immunoprecipitation.

 Ordering Information

 

RNA Isolation	DNA Isolation	Protein Isolation
Chloroform Isopropyl alcohol 75% ethanol (in DEPC-treated water) RNase-free water or 0.5% SDS Centrifuge and rotor capable of reaching up to 13000 RPM Polypropylene microcentrifuge tubes Water bath or heat block (55-60°C)	Chloroform 100% ethanol 75% ethanol 0.1 M sodium citrate in 10% ethanol 8 mMNaOH Centrifuge and rotor capable of reaching up to 13000 RPM Polypropylene microcentrifuge tubes	Chloroform Isopropyl alcohol 100% ethanol 0.3 M Guanidine hydrochloride in 95% ethanol 1% SDS Centrifuge and rotor capable of reaching up to 13000 RPM Polypropylene microcentrifuge tubes

Advantages of using TRIZIN for RNA Isolation

1. TRIZIN enables the isolation of RNA by a single step color guided biphasic extraction method from biological samples of different sources (e.g., human, other animals, plants, yeast, bacterial and virus origin, etc.).
2. The isolated RNA is substantially free of genomic DNA and hence ready for Reverse Transcriptase PCR reaction. The bright purple color of the reagent makes the phase separation (purple colored organic phase and colorless aqueous phase) clearly visible.
3. Color guided separation not only minimizes the chance of getting contamination during the collection of aqueous phase but also offers a substantial yield in RNA isolation.

Homogenization

Determine your sample type and perform homogenization 4˚C according below mentioned table-1. The sample volume should not exceed 10% of the volume of TRIZIN Reagent used for homogenization. Be sure to use the indicated amount of TRIZIN Reagent, because an insufficient volume can result in DNA contamination of isolated RNA. Homogenized samples can be stored at –60 C to –70 °C for at least one month

Phase separation

1. Incubate the homogenized sample (see Homogenization) for 5 minutes at room temperature to permit complete dissociation of the nucleoprotein complex.
2. Add 0.2 mL of chloroform per 1 mL of TRIZIN Reagent used for homogenization. Cap the tube securely.
3. Shake tube vigorously by hand for 15 seconds.
4. Incubate for 2–3 minutes at room temperature and repeat the step 3 for at least two times.
5. Centrifuge the sample at 12,000 × g for 15 minutes at 4°C.
6. Remove the aqueous phase of the sample by angling the tube at 45° and pipetting the solution out. Avoid drawing any of the inter phase or organic layer into the pipette when removing the aqueous phase.
7. Place the aqueous phase into a new tube and proceed to RNA Isolation Procedure, for save the inter phase and organic phenol-chloroform phase for DNA and Protein Isolation.