Found 1 discussion for this experiment
4 years ago
4 years ago by Ralf Friedmann
I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?
Found 3 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.
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