Found 2 discussions for this experiment
1 year ago
1 year 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?
1 year ago
1 year 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 2 matching solutions for this experiment
|- Complete disruption of plasma membranes of cells and organelles is absolutely required to release all the nucleic acids contained in the sample.
- Different samples require different methods to achieve complete disruption.
- Incomplete disruption results in significantly reduced nucleic acid yields.
- Overloading the spin columns significantly reduces nucleic acid yields.
|- Homogenizing the material is necessary to redice the viscosity of the lysates caused by cell disruption.
- Incomplete homogenization results in inefficient binding of DNA and RNA and therefore significantly reduced yield and purity of nucleic acids.
- Excessive homogenization, on the other hand, results in shorter genomic DNA fragments.
- The centrifugation temperature should be 20–25ºC.
- Warm the lysate to 37ºC before transferring it to the AllPrep DNA spin column.