1. If samples are frozen ensure that they have thawed completely before starting this procedure.
Add 90μl of sample to 150μl of Reagent 1 (or scale up for larger sample volume).
Mix thoroughly using a pipette or vortex mixing.
2. Incubate for one minute at room temperature.
At this point, DNA is stabilized for 90 days and RNA is stabilized for up to 7 days at room temperature, provided there is no further processing.
3. Take 5μl of the above-lysed mixture and combine it with 20μl of Reagent 2 (or scale up for larger sample volumes maintaining the 1:4 ratio).
Once processed with Reagent 2, samples should be used within 4 hours or frozen at -20 ̊C.
4. Add appropriate volume into PCR master mix (e.g. 5μl per 25μl reaction) or continue directly to other downstream techniques.
Dilute Sample Protocol
1. When handling very dilute samples such as saliva the ratio of sample to Arcis Reagent 1 can be increased to 1:1 to avoid further dilution (90μl of sample to 90μl of Reagent 1).
2. Samples that have been processed as in step 1 of the standard protocol can be added to Reagent 2 at 1:3, 1:2, or 1:1 ratio to reduce sample dilution.
|Extract from lysis reaction (μl)
||Reagent 2 (μl)