QIAGEN Inc
RNeasy Mini Kit
74104
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I am interested in the expression level of several inflammatory genes in peripheral blood mononuclear cells. I used this kit for RNA cleanup after extraction with Trizol. This kit was recommended by several collaborators and it worked well for me.
For RNA cleanup
Peripheral blood mononuclear cell
1. Add 4 volumes of ethanol (96%-100%) to Buffer RPE for a working solution. 2. Adjust to a volume of 100 ul with water. Then add 350 ul Buffer RLT, and gently vortex. 3. Add 250 uL of ethanol, mix by pipetting. Do not centrifuge. Proceed immediately to next step. 4. Load 700 uL the mixture onto the column placed on a 2 mL collection, centrifuge at 8,000 g for 15 S. Dump the flow-through fluid. 5. Add 500 uL of RPE buffer onto column, close the lid, centrifuge at 8,000 g for 15 S to wash the membrane. Discard the flow-through. 6. Add another 500 uL of RPE buffer, centrifuge at 8,000 g for 2 mins. 7. Place column in a new 2 ml collection tube. Centrifuge at 12 000 g for 1 min. 8. Transfer the column to a 1.5 ml collection tube; add 30 uL of RNAase-free water. Centrifuge at 8,000 g for 1 min to elute the RNA.
Take extra caution to avoid RNase contamination by setting up a special area for the work. Always use new gloves.
High quality RNA can be obtained by this kit.
DNAse on column digestion is recommended to eliminate genomic DNA contamination. The DNase kit needs to be purchased separately.
Instruction is easy to follow. High quality RNA can be obtained.
Pricey compared with similar products.
If the quality of the RNA is critical for downstream applications, I would recommend this kit.
Product Name: RNeasy Mini Kit (50)Supplier Page