The Epigenetics Company

ZR-Duet™ DNA/RNA MiniPrep

Quick isolation and separation of genomic DNA and total RNA (up to ~25 μg each) from a wide range of sources using Fast-Spin column technology.
DNA/RNA products are suitable for use in PCR, RT-PCR, and other procedures.
Omits the use of organic denaturants and proteases.

Product Name Size Catalog # Price Qty
ZR-Duet™ DNA/RNA MiniPrep 50 Preps. D7001
$297.00

About ZR-Duet™ DNA/RNA MiniPrep

The ZR-Duet™ DNA/RNA MiniPrep provides a quick method for parallel purification of high quality genomic DNA and total RNA from small amounts of cells and tissue. The kit isolates both genomic DNA and large and small RNA species without the use of phenol or reducing agents. Small RNAs (e.g., tRNAs, microRNAs) can be recovered following a simple adjustment of the RNA isolation protocol no extra steps are required! Both DNA and RNA (up to ~25 µg each) from 102 to 107 cells can be isolated in less than 15 minutes.


Format Spin Column
Processing Time 15 min
Equipment Microcentrifuge
Sample Sources Cells, small amounts of easy-to-lyse tissue, buffy coat, buccal cells, plasma, serum, and other biological liquids. Not compatible with whole blood. (For purification of DNA from whole blood see the Quick-gDNA™ MiniPrep (D3024) and for RNA see the ZR Whole-Blood RNA MiniPrep (R1020))
Storage -70°C
Sample Size 102 to 5x106 cells in suspension or as tissue.
Purity High quality genomic DNA and total RNA (A260/A280 > 1.8, A260/A230 > 1.8) is recovered.
Size Limit Capable of recovering genomic DNA up to and above 40 kb. In most instances, mitochondrial DNA and viral DNA (if present) will also be recovered. Total RNA ≥ 17 nucleotides can be recovered.
Recovery Up to 25 μg DNA/RNA can be eluted into ≥ 25 μl

DNA and RNA purified using the ZR-Duet™ DNA/RNA MiniPrep. Genomic DNA (lane 1, 2) and total RNA (lane 3, 4) isolated from human epithelial cells (HCT 116) with the ZR-Duet™ DNA/RNA MiniPrep. M is a 1 kb DNA Marker (Zymo Research).
 

PCR amplification of ß-actin transcript (353 bp fragment shown) following DNA and RNA isolation from human epithelial cells (HCT 116) with the ZR-Duet™ DNA/RNA MiniPrep: PCR positive control (DNA template; lane 1, 2, 3), PCR negative control (RNA template; lane 4, 5, 6), RT-PCR (lane 7, 8, 9). M1 and M2 are 1 kb and 100 bp DNA Markers, respectively (Zymo Research).

  1. Sample Preparation
    1. Adherent Cells:  Cells can be lysed directly in the culture container by removing liquid medium and adding DNA/RNA Lysis Buffer1 directly to the monolayer (e.g., 400 µl for 102 to 5x106 cells).  Remove cells from the culture surface by pipetting, scraping, etc.  Proceed to Step 2.
    2. Cells in Suspension:  Pellet the cells by gentle centrifugation (e.g., 5 minutes at 500 x g).  Remove the supernatant completely and resuspend the cell pellet in 400 µl DNA/RNA Lysis Buffer1.  Vortex briefly. Proceed to Step 2.
    3. Solid Tissue Samples:  Add 400 µl DNA/RNA Lysis Buffer1 to fresh or frozen tissue (up to ~25 mg) and homogenize the sample (e.g., using a Dounce or similar homogenizer).  Proceed to Step 2.
    4. Liquid Samples:  Add 3 volumes of DNA/RNA Lysis Buffer1 for every volume of sample (e.g., 300 µl buffer to 100 µl sample).  Proceed to Step 2.
  2. Transfer the sample from Step 1 into a Zymo-Spin™ IIIC Column2,3 in the Collection Tube and centrifuge at ≥12,000 x g for 1 minute. Save the flow-through for RNA and the column for DNA purification!


DNA Purification

  1. Transfer the Zymo-Spin™ IIIC Column into a new Collection Tube.
  2. Add 200 µl DNA Prep Buffer to the column and centrifuge at ≥12,000 x g for 30 seconds.
  3. Add 200 µlDNA Pre-Wash Buffer to the column and centrifuge at ≥12,000 x g for 1 minute. Discard the flow-through.
  4. Add 500 µl g-DNA Wash Buffer to the column and centrifuge at ≥12,000 x g for 30 seconds. Discard the flow-through.
  5. Centrifuge the Zymo-Spin™ IIIC Column in an emptied Collection Tube at ≥12,000 x g for 2 minutes.  Remove the Zymo-Spin™ IIIC Column carefully from the Collection Tube and transfer it into a clean microcentrifuge tube.
  6. Add ≥50 µl DNase/RNase-Free Water directly to the column matrix and let stand 2 to 5 minutes at room temperature, then centrifuge at top speed for 30 seconds.  The eluted DNA can be used immediately or stored at ≤-20ºC.


RNA Purification

  1. Add 0.8 volume5 ethanol (95-100%) to the flow-through in the Collection Tube6 from Step 2 (e.g., 320 µl ethanol to 400 µl flow-through) and mix well by pipetting.
  2. Transfer the sample from Step 3 into a Zymo-Spin™ IIC Column3,4 in a Collection Tube and centrifuge at ≥12,000 x g for 1 minute.  Discard the flow-through.
  3. Add 400 µl RNA Prep Buffer to the column and centrifuge at ≥12,000 x g for 1 minute.  Discard the flow-through.
  4. Add 700 µl RNA Wash Buffer to the column and centrifuge at ≥12,000 x g for 30 seconds.  Discard the flow-through. Repeat the wash step with 400 µl RNA Wash Buffer.
  5. Centrifuge the Zymo-Spin™ IIC Column in an emptied Collection Tube at ≥12,000 x g for 2 minutes.  Remove the Zymo-Spin™ IIC Column carefully from the Collection Tube and transfer it into an RNase-free tube.
  6. Add ≥25 µl DNase/RNase-Free Water directly to the column matrix and let stand 1 minute at room temperature.  Centrifuge at 10,000 x g for 30 seconds. The eluted RNA can be used immediately or stored at ≤-70°C.
For specific notes and additional information, please see the product protocol PDF.
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