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 3 matching solutions for this experiment
|- On-column digestion is preferable
- For high gDNA elimination spin at 6000rpm for 5 min instead of the quick high speed spin
|- 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.
|- If the lysate viscous than normal. Pass these cell or tissue lysates several times through a 21-gauge needle. If the yield of mRNA is low Increase the ratio of beads to Lysis/Binding Buffer or add RNase-free Proteinase K to the sample lysate directly before the 10 minute binding incubation
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