The QIAamp RNA Blood Mini Kit is designed for the extraction of total RNA from human whole blood and other samples. The protocol is pre-optimized for different sample types, including whole blood, cultured cells (e.g. NIH3T3, HeLa, COS-7, Huh-7) and tissue (e.g. brain, heart, kidney, liver, spleen and lung). The extraction is column-based (microspin technology). What I like best about this kit is that it does not involve any toxic substances, such as phenol/chloroform, or gradient ultracentrifugation.
Extraction of RNA from blood involves steps where erythrocytes are selectively lysed and leukocytes are recovered by centrifugation. The leukocytes are then lysed using highly denaturing conditions that immediately inactivate RNases, allowing the isolation of intact RNA. After homogenization of the lysate by a brief centrifugation through a QIAshredder spin column, ethanol is added to adjust binding conditions and the sample is applied to the QIAamp spin column. RNA is bound to the silica membrane during a brief centrifugation step. Contaminants are washed away and total RNA is eluted in 30 ìl or more of RNase-free water for direct use in any downstream applications.
I have used this kit successfully over the last couple of years to isolate total RNA from human whole blood. Most importantly, this kit is compatible with all anticoagulants, in which blood can be collected, including EDTA, citrate, heparin or ACD, whereas other manufacturers generally recommend only EDTA. The volume of blood can be easily scaled up or down as per leukocyte count (typically 4000-7000 leukocytes per microliter). The average yield we obtain is somewhere 2-2.5 ug in 30 ul from 1.5 ml of whole blood at 4000-7000 leukocyte per microliter (for elevated leukocyte count, less then 1.5 ml of blood must be used). We require only 1 ug for our downstream applications, like quantitative PCR. We check the quality and quantity of total RNA by using spectrophotometer (NanoDrop).
As Qiagen says – Qiagen guarantees the performance of all products in the manner described in our product literature. They really mean that. I have been using this kit for pretty long time and I can really guarantee its performance and resulting satisfaction.
QIAamp RNA Blood Mini Kit From Qiagen
It is an easy and efficient method for extracting total RNA from different biological samples.
The Bottom Line
A reliable kit for quantitative determination for gene expression assays.