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Quick-RNA™ MidiPrep

Quick recovery of RNA from a wide range of cell types and tissue using Fast-Spin column technology.
Column and plate designs allow for elution of highly concentrated RNA.
Omits the use of organic denaturants and proteases.

Product Size Catalog # Price Qty
Quick-RNA™ MidiPrep 25 Preps. R1056

About Quick-RNA™ MidiPrep

The Quick-RNA™ MidiPrep is an innovative product designed for the easy, reliable, and rapid isolation of total RNA from cultured cells or solid tissue samples. The procedure combines a unique, single-step RNA extraction/binding buffer with Fast-Spin column technology to yield high quality RNA in minutes. The method is easy: simply add the provided ZR RNA Buffer to extract total RNA from the cells of interest and then purify the RNA using the provided spin column. The result is highly-concentrated, purified RNA that is suitable for subsequent RNA-based methods including RT-PCR, hybridization, etc.

Format Spin column.
Processing Time 15 min
Sample Sources Cells from culture or tissue samples, buffy coat, plasma, serum, and other biological liquids. Not compatible with whole blood. (For purification of RNA from whole blood see the ZR Whole-Blood RNA MiniPrep (R1020))
RNA Purity High quality RNA (A260/A280 > 1.8, A260/A230 > 1.8) suitable for all downstream RNA-based manipulations.
RNA Recovery Up to 1 mg RNA can be eluted into ≥ 200 µl RNase-free water allowing for a highly concentrated sample.
Sample Size ~103 to 108 cells.
RNA Storage -70ºC
Equipment Needed Vacuum manifold, microcentrifuge.

Human RNA isolated from cell culture with the Quick-RNA™ MiniPrep Kit. M: 1 kb DNA marker (Zymo Research); 1, 2: duplicate RNA preparations.

PCR amplification of a ß-actin transcript (353 bp fragment shown) following RNA isolation from human epithelial cells (HCT 116) with the Quick-RNA™ MidiPrep: RT-PCR (lanes 1, 2), PCR negative control (RNA template; lanes 3, 4). M1 and M2 are 100 bp and 1 kb DNA Markers, respectively (Zymo Research).

Step 1

Sample Homogenization and Cell Lysis:  Follow an applicable method (A-D).

A. Adherent Cells: Cells can be lysed directly in the culture container by removing liquid medium and adding ZR RNA Buffer1 directly to the monolayer (e.g., 3 ml  for 5x106 cells).  Remove cells from culture surface by pipetting, scraping, etc.

B. Cells in Suspension: Pellet cells by gentle centrifugation (e.g., 5 minutes at 500 x g).  Remove the supernatant completely and resuspend the cell pellet in 3 ml ZR RNA Buffer1. Vortex briefly.

C. Solid Tissue Samples: Add 3 ml ZR RNA Buffer1 to fresh or frozen tissue (up to ~100 mg) and homogenize the sample (e.g., using a Dounce or similar homogenizer)Important!  To eliminate tissue debris, if present, sample lysates may be spun at ≤500 x g for 1 minute.

D. Liquid Samples: Add 3 volumes of ZR RNA Buffer1 to every volume of sample (e.g., 3 ml of buffer to 1 ml sample).

For total RNA including small RNAs

For total RNA without small RNAs

Add 1 volume ethanol (95-100%) to the lysate and mix thoroughly by pipetting.    Go directly to Step 2.
Step 2

Transfer lysate into the Zymo-Spin™ V-E Column with Reservoir mounted on a vacuum manifold and start vacuum4.

Step 3

Remove the reservoir and transfer the Zymo-Spin™ V-E Column into a Collection Tube.  Centrifuge the column at ≥12,000 x g for 30 seconds.

Step 4

Add 400 µl RNA Pre-Wash Buffer to the column and centrifuge at ≥12,000 x g for 1 minute.  Discard the flow-through.

Step 5

Add 400 µl RNA Wash Buffer to the column and centrifuge at ≥12,000 x g for 30 seconds.  Discard the flow-through and repeat Step 5.

Step 6

Centrifuge the Zymo-Spin™ V-E Column at ≥12,000 x g for 2 minutes in the emptied Collection Tube to ensure complete removal of the wash buffer.

Step 7

Place the Zymo-Spin™ V-E Column into an RNase-free tube.  Add ≥200 µl DNase/RNase-Free Water5 directly to the column matrix and let stand at room temperature for 1 minute.  Centrifuge at top speed for 1 minute.  Eluted RNA can be used immediately or stored at ≤-70 ºC.

For specific notes and additional information, please see the product protocol PDF.
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