SV Total RNA Isolation System

RNA isolation / purification Bacteria - Gram positive Streptococcus pneumoniae

Experiment
RNA isolation / purification Bacteria - Gram positive Streptococcus pneumoniae
Product
SV Total RNA Isolation System from Promega
Manufacturer
Promega

Protocol tips

Protocol tips
Genomic DNA and total RNA were isolated from S. pneumoniae strains using a Wizard genomic DNA isolation kit and SV total RNA isolation system (Promega), respectively, following the manufacturer's instructions, except that cells were incubated with 0.1% deoxycholic acid (Sigma) at 37°C for 10 min before extraction. RNasin (0.5%; Promega) was added to extracted RNA to prevent it from degradation. cDNA was derived and amplified from RNA using an Access reverse transcription-PCR (RT-PCR) system (Promega) and target-specific primers.

Publication protocol

Genomic DNA and total RNA were isolated from S. pneumoniae strains using a Wizard genomic DNA isolation kit and SV total RNA isolation system (Promega), respectively, following the manufacturer's instructions, except that cells were incubated with 0.1% deoxycholic acid (Sigma) at 37°C for 10 min before extraction. RNasin (0.5%; Promega) was added to extracted RNA to prevent it from degradation. cDNA was derived and amplified from RNA using an Access reverse transcription-PCR (RT-PCR) system (Promega) and target-specific primers.



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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

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Manufacturer protocol

Download the product protocol from Promega for SV Total RNA Isolation System below.

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