TRIzol Reagent - Invitrogen

Slower speed for centrifugation: Table-top centrifuges that can attain a maximum of 2600 x g can be used if the
centrifugation time is increased to 30-60 min.
RNA Wash
(back to Table of Contents)
(back to Protocol and Application Notes)
(back to RNA Isolation with TRIzol LS)
Remove the supernatant. Wash the RNA pellet once with 75% ethanol, adding at least 1 ml of 75% ethanol per 0.75 ml of TRIzol
LS Reagent used for the initial homogenization. Mix the sample by vortexing and centrifuge at no more than 7500 X g for 5 min.
at 4ºC.
Possible stopping point: The RNA precipitate can be stored in 75% ethanol at 4ºC for at least one week, or at least
one year at -20ºC.
Redissolving the RNA Pellet
(back to Table of Contents)
(back to Protocol and Application Notes)
(back to RNA Isolation with TRIzol LS)
Briefly dry the RNA pellet (air-dry preferred or vacuum-dry for 5min.). Dissolve RNA in RNase-free water or 0.5% SDS solution
by passing the solution a few times through a pipette tip, and incubating for 10 min. at 55-60ºC.
Overdrying the RNA pellet: Air drying the RNA pellet is preferred over drying by centrifugation under vacuum. It
is important not to let the RNA pellet dry completely as this will greatly decrease its solubility.
Additional Notes for RNA Isolation with TRIzol LS
RNAses
(back to Table of Contents)
(back to Protocol and Application Notes)
(back to RNA Isolation with TRIzol LS)
In the presence of TRIzol LS Reagent, RNA is protected from RNase contamination.
Tubes to use with TRIzol LS
(back to Table of Contents)
(back to Protocol and Application Notes)
(back to RNA Isolation with TRIzol LS)
See section about “Tubes to use with TRIzol”.
Troubleshooting RNA Isolation with TRIzol LS
For additional information see Troubleshooting RNA Isolation with TRIzol Notes Section above.
Low Yield of RNA
(back to Table of Contents)
(back to Protocol and Application Notes)
(back to RNA Isolation with TRIzol LS)
Incomplete homogenization or lysis of samples.
Final RNA pellet incompletely redissolved.
OD reading may vary due to solution the sample is stored in AND what it was diluted in prior to quantitation.
Wilfinger, et. al., in (1997) BioTechniques 22.3, 478 report that increases in ionic strength may significantly reduce
nucleic acid absorbance, thereby affecting quantitative determinations based on A260.
26