1. Isolation of RNA from small quantities of tissue (1 to 10 mg) or Cell (102 to 104) Samples: Add 800 µl of TRIZOL to the tissue or cells. Following
sample lysis, add chloroform and proceed with the phase separation as
described in step 2. Prior to precipitating the RNA with isopropyl alcohol,
add 5-10 •Ïg RNase-free glycogen (Cat. No 10814) as carrier to the
aqueous phase. To reduce viscosity, shear the genomic
2. DNA with 2 passes through a 26 gauge needle prior to chloroform
addition.
3. The glycogen remains in the aqueous phase and is co-precipitated with the
RNA. It does not inhibit first-strand synthesis at concentrations up to 4
mg/ml and does not inhibit PCR.
4. After homogenization and before addition of chloroform, samples can be
stored at -60 to -70°C for at least one month. The RNA precipitate (step 4,
RNA WASH) can be stored in 75% ethanol at 2 to 8°C for at least one
week, or at least one year at –5 to -20°C.
5. Table-top centrifuges that can attain a maximum of 2,600xg are suitable
for use in these protocols if the centrifugation time is increased to 30-60
minutes in steps 2 and 3.
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