Found 2 discussions for this experiment
4 years ago
4 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
|- For RNA to dissolve better and to yeild high levels, preheat elution buffer at 95C.
- Addition of β-mercaptoethanol in lysis buffer can be skipped because the phenol will do the job of nuclease inactivation
- To capture more amount of mRNA, modify the amount of lysis buffer :ethanol (from 1:1 to 1:1.5) during the binding step
|- Do vortex for 2-plus minutes during Acid-Phenol:Chloroform extraction step. This will facilitate the removal of tightly bound proteins typically associated with RNA
|- 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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