Found 2 discussions for this experiment
3 years ago
3 years ago by Paul G. Macon
How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?
4 years ago
4 years ago by Aaron Stege
I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?
Found 7 matching solutions for this experiment
|- To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).|
|- Include DNAse treatment for 15-20min.
- Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C.
- Use water to elute the RNA that is warmed to ~60`C.
|- When processing <2 µg tissue, carrier RNA may be added to the lysate. Foaming can be reduced by adding Reagent DX at a final concentration of 0.5% (v/v) before disruption and homogenization.
- Add 4 volumes of ethanol (96–100%) to Buffer RPE for a working solution.
|- To allow complete penetration by formalin, use tissue samples less than 5 mm thick
|- Perform all centrifugation steps using a microcentrifuge placed at 15–25°C
|- Do not exceed fixation time of 24 hours as it results in poor performance in downstream assay
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