TRIzol™ LS Reagent

RNA isolation / purification Bacteria - Gram positive Listeria monocytogens

Experiment
RNA isolation / purification Bacteria - Gram positive Listeria monocytogens
Product
TRIzol™ LS Reagent from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Upstream tips
TRIzol LS reagent is intended for liquid samples, using solid samples will decrease RNA yield

Publication protocol

For RNA isolation, pellets were resuspended in 750 µl TRIzol Reagent (Life Technologies). Cells were disrupted using beadbeating in Lysing Matrix A tubes (MP Biomedicals) with a FastPrep FP120 instrument (MP Biomedicals) with the following parameters: three times 45s at speed 5.5 at 4°C. RNA isolation was performed according to the manufacturer’s instructions. Concentration of nucleic acids was measured with a Nanodrop instrument (Shimadzu BioSpec-nano) and remaining DNA was removed using the Turbo DNA-free Kit (Life Technologies) according to manufacturer’s instructions. The absence of DNA was confirmed by performing a PCR using qacH fwd and rev primers. PCR conditions were the same as indicated above, except that 40 cycles were used.

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Discussion

Discussion

5 years ago

Author: Paul G. Macon United States

Some help with RNA isolation using Trizol

Hello! I used Trizol to extract total RNA from in-vitro cultured bacteria (1 X 10^8 cells). After phase separation, I mixed ~0.4 ml of the upper phase which contains RNA with 0.5 mL cold isopropanol. However, the amount of RNA when measured in Nanodrop was very low. In addition, the ratio between 260 and 230 was around 0.1 to 0.5. Is there a chance that my sample was contaminated by the Trizol reagent? When I collected the aqueous phase I made sure to not touch the lower phase. What should I do?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Bacteria - Gram positive Listeria monocytogens using TRIzol™ LS Reagent from Thermo Fisher Scientific.

Paper title
Tn - A Novel Transposon in Responsible for Tolerance to Benzalkonium Chloride
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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for TRIzol™ LS Reagent below.

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Videos

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