QIAGEN - Sample & Assay Technologies
RNeasy Micro Kit (50)
74004
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This kit works well for any small amount RNA isolation. We can get high quality RNA from 100,000 T cells using this kit. The instructions are simple and easy to follow. Most optimization guidelines are detailed in the back of the Qiagen handbook making troubleshooting a breeze. Well worth the cost for quality results of small or sensitive samples.
RNA Isolation
Less than 1,000,000 murine cells
After collecting cells, lyse cells in RLT Buffer, pipette cells in buffer directly into QIAshredder and centrifuge for 2 minutes (or lyse cells in a fine needle at least 10 times). Remove shredder and add 1 volume of 70% ethanol and mix well. then transfer to a MinElute spin column in a collection tube and centrifuge for 15 sec. Discard flow-through and add RW1 buffer to the spin column and centrifuge 15 sec. Discard flow-through and add DNase mixed at a 1 to 7 dilution in RDD Buffer and incubate 15 min on benchtop. Add Buffer RW1 and centrifuge 15 sec. Using a new collection tube add RPE Buffer and centrifuge 15 sec. Discard flow-through and add 80% ethanol, centrifuge 2 min. Discard flow through and centrifuge again with an open lid for 5 min. Then elute RNA in 14ul of water. Place water directly on white membrane to collect all of the RNA and centrifuge for 1 min.
Perform all steps even if they are indicated as optional.
High quality RNA for use in downstream analysis.
N/A
Consistent and reliable results for great RNA
If using DNase it comes in a separate kit.
Kits can always be pricey but this one works for the cost and is comparable if not better than other kits for RNA quality.