Max Bacterial Enhancement Reagent

Yes you can. Here is a reference to a paper, as well as a brief description of method from the paper.

Genes to Cells (2001) 6:121-129 (under the heading 'RNA isolation and RT-PCR')

TRIzol LS Reagent (LifeSciences) was used according to the manufacturer's instructions to extract total RNA from sucrose gradient fractions. Briefly, 250 mL of each fraction was added to 750 mL TRIzol LS Reagent and shaken vigorously for 15 s. After a 10-min incubation at room temperature, 150 mL chloroform was added, followed by vigorous shaking and brief incubation at room temperature. Samples were then spun at 14,000 g for 10 min in a tabletop microcentrifuge. Five micrograms of nuclease-free glycogen were added to 300 mL of the aqueous phase and nucleic acids were precipitated with the addition of an equal volume of 2-propanol. After centrifugation at 14,000 g for 30 min at room temperature, the pellet was washed once with 75% ethanol and resuspended in 20 mL of nuclease-free, sterile water. Five microlitres of total RNA were used as substrate for random-primed cDNA synthesis using Superscript II modified MMLV reverse transcriptase (Gibco/Life Sciences).

Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.

TRIzol Reagent contains phenol and guanidine isothiocyanate, which allow for isolation of nucleic acids from proteins. Further partitioning of the nucleic acids occurs in a pH dependent manner. At pH 7.0 or higher, DNA and RNA partition into the aqueous phase. At an acidic pH, below 7.0, DNA is denatured and will move into the organic phase (interphase) yet the RNA remains in the aqueous phase.TRIzol Reagent has pH about 5.

Blood collected with EDTA typically has the highest DNA contamination, blood collected with heparin typically has less than that collected with EDTA, and blood collected with citrate shows the least DNA contamination of the three. (Formulation for citrate solution: 3.8% (w/v) which is 3.8 g/100 mL of water. Use 0.5 mL for every 4.5 mL of blood. Rock gently back and forth after adding citrate solution to mix.) Adding 12 µL of 5 N acetic acid per milliliter of TRIzol Reagent may help, although there may still be a problem with DNA contamination. Using plasma or serum works best. The fresher the blood sample the better the RNA. Degraded RNA has been observed in blood that has been processed in as little as two hours after drawing.

Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.

There are a couple of reasons that the aqueous phase may appear pinkish. It may be due to the tissue you are using. This is common with skin samples. It is assumed that there is fat in these samples, and the fat micelles “try to spin to the top of the tube” during the centrifugation but are unable to get there. In skin samples, the micelles pick up melanin pigment and cause the aqueous phase to appear colored. Fat micelles may also pick up pigment from the TRIzol Reagent itself, resulting in a pinkish color. If a sample is thought to contain fat, the sample homogenate in TRIzol Reagent may be centrifuged prior to addition of chloroform. The fat will appear as a clear layer at the top of the supernatant; this should be pipetted off and discarded.

In addition, if a sample contains a lot of blood, the aqueous phase may appear cloudy and/or yellowish (this may be due to iron in the hemoglobin coming out).

If the centrifuge used is not cold, the organic phase will be a deeper maroon color; some of this color may come into the aqueous phase and cause it to appear orange or yellow.

Alternatively, a pinkish aqueous phase may also be caused by over-dilution of the sample (i.e., the sample:TRIzol Reagent ratio which is greater than 1:10), as well as too much salt or protein in the sample. This can cause premature phase separation, which can be remedied by adding a bit more TRIzol Reagent to the sample. If the RNA is isolated from a pinkish aqueous phase, chances are that it will be contaminated with DNA. Although this should not be significant as TRIzol Reagent is formulated to prevent premature phase separation.

Find additional tips, troubleshooting help, and resources within our RNA Sample Collection, Protection, and Isolation Support Center.

TRIzol LS Reagent is a complete, ready-to use reagent for easy and simultaneous isolation of total RNA, DNA, and proteins from liquid samples. The reagent, a mono-phasic solution of phenol and guanidinium thiocyanate, is an improvement to the single-step RNA isolation method developed by Chomzcynski and Sacchi. TRIzol LS Reagent is similar to the original TRIzol Reagent in composition and results of use. TRIzol LS Reagent is designed for use with liquid samples such as blood and virus preparations in which large volumes of aqueous samples need to be processed, while TRIzol Reagent is designed for cell cultures or tissues. It is formulated to accommodate processing of more liquid sample per unit of reagent compared to the original formula.

The only difference between TRIzol Reagent and TRIzol LS Reagent is the concentration of components. TRIzol  LS Reagent is slightly more concentrated. The formula allows lower quantities of reagent to be used relative to a liquid sample. (TRIzol = 10:1 required, TRIzol LS = 3:1 required). If you buy TRIzol LS Reagent, but want to use it like TRIzol Reagent (on solid samples), there will probably be a decrease in yield vs. using regular TRIzol Reagent. TRIzol LS Reagent should NOT be used undiluted with solid samples. To dilute: take 750 µL TRIzol LS Reagent + (50 to 100 mg tissue + water to make 250 µL).