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Quick-cfDNA™ Serum & Plasma Kit

High-quality DNA, including cell-free, is easily and robustly purified from up to 10 ml of serum, plasma, amniotic fluid or cerebrospinal fluid.
Zymo-Spin™ technology enables elution of DNA in as little as 35 μl and ensures it is ready for all sensitive downstream applications such as qPCR and Next-Generation sequencing.

Product Size Catalog # Price Qty
Quick-cfDNA™ Serum & Plasma Kit 50 preps D4076

About Quick-cfDNA™ Serum & Plasma Kit

Quick-cfDNA™ Serum & Plasma Kit provides a simple and reliable method for the rapid preparation of high-quality circulating cell-free DNA from serum, plasma, amniotic fluid, cerebrospinal fluid (CSF), and saliva. A combination of chemical and enzymatic methods are used to efficiently recover total DNA (including cell-free apoptotic, necrotic, mitochondrial and viral DNA) linearly from up to 10 ml of sample (Reference Figure A). Zymo-Spin technology allows for ultra-pure DNA to be eluted in as little as 35 μl water. The resulting DNA is suitable for all subsequent analyses and molecular manipulations such as qPCR, Next-Generation sequencing and DNA methylation analyses. Zymo’s serum and plasma cell-free DNA extraction technology will empower your discovery of circulating DNA biomarkers.

Learn more about cell-free DNA: 

Circulating cell-free DNA is found in various body fluids, including blood, and is generally derived from apoptotic cells, necrotic cells, and intact cells that were released into the bloodstream and eventually lysed [1-2]. Circulating cell-free DNA in serum and plasma is usually composed primarily of cell-free DNA fragments derived from healthy cells, however, in cancer patients circulating tumor DNA can be detected with a higher signal to noise ratio than whole blood for non-invasive diagnostics.  Therefore, cell-free tumor DNA shed from circulating tumor cells is being investigated as a source of biomarkers for the early diagnosis, prognosis and monitoring of cancer therapy [3]. The utility and value of serum and plasma for non-invasive molecular diagnostics is demonstrated by the use of cell-free fetal DNA for sex determination and prenatal diagnosis of fetal chromosomal aneuploidy and other genetic disorders [4].

DNA Recovery Recover DNA ≥ 100bp (optimized for recovery of cell-free DNA). Yields can vary considerably among different individuals. Typically DNA recovery ranges from 1-100 ng/ml of plasma or serum. The yield varies depending on the sample source and the health of the donor.
Equipment Water bath or heat block (55ºC), microcentrifuge, vacuum/vacuum manifold or swinging bucket centrifuge.
Sample Sources Serum, plasma, amniotic fluid, and cerebrospinal fluid (CSF).
DNA Purity High-quality DNA is ready for all sensitive downstream applications such as qPCR and Next-Generation sequencing.
Storage Eluted DNA should be stored at ≤ -20ºC.
Recovery Volume ≥ 35 μl of DNA Elution Buffer or DNase free water.
Processing Volume Plasma – Single centrifugation: up to 3 ml
Plasma – Double centrifugation: up to 10 ml
Serum/Amniotic fluid/Cerebrospinal fluid (CSF) – up to 10 ml
Saliva – up to 1 ml
Cell-free Saliva – up to 5 ml

Cell-free DNA recovery is directly scalable using the Quick-cfDNA™ Serum & Plasma Kit. (A) Graphs and (B) gel image show the linear recovery of cfDNA from human plasma and serum (healthy female donors), as measured by Tapestation 2200 (Agilent, in duplicates).

Total DNA is efficiently purified from cell-free biological fluids with the Quick-cfDNA™ Serum & Plasma Kit. Total DNA, including both high and low molecular weight species, purified (duplicates) from human maternal plasma, amniotic fluid and cerebrospinal fluid was analyzed by Tapestation 2200 (Agilent).

[1] Schwarzenbach H, Hoon DS, Pantel K. Nat. Rev. Cancer. 2011, 11: 426-437.
[2] Jahr S, Hentze H, Englisch S, et al. Cancer. Res. 2001, 61: 1659-1665.
[3] Schwarzenbach H, Alix-Panabières C, Muller I, et al. Clin Cancer Res. 2009, 15: 1032–1038.
[4] Lewis C, Hill M, Skirton H, Chitty LS. Eur. J. Hum. Genet. 2012, 20: 1127–1133.

Reagent Preparation:
  • Prior to use, add 6.5 ml of Proteinase K Storage Buffer to each Proteinase K (125 mg) tube. The final concentration of Proteinase K is ~20 mg/ml. Store at -20ºC after mixing.
  • Prior to use, add 48 ml 95-100% ethanol to the 12 ml S&P DNA Wash Buffer.

  1. Add 50 µl of S&P 5X Digestion Buffer to every 200 µl of serum, plasma or biological fluid and mix thoroughly (Reference Table 1).

    Note: For an input other than 200 µl of Biological Fluid, adjust S&P 5X Digestion Buffer, Proteinase K, and S&P DNA Binding Buffer proportionally.
  2. Add 20 µl of Proteinase K to every 200 µl of serum, plasma or biological fluid and mix thoroughly (Reference Table 1).
  3. Incubate at 55ºC for 30 minutes.
  4. Add two volumes of S&P DNA Binding Buffer to the digested sample and mix thoroughly (Reference Table 1).

Sample Volume 200µl 1 ml 3 ml 5 ml 10 ml
S&P 5X Digestion Buffer 50 µl 250 µl 750 µl 1.25 ml 2.5 ml
Proteinase K 20 µl 100 µl 300 µl 500 µl 1 ml
Mix thoroughly and incubate at 55ºC for 30 minutes.
S&P DNA Binding Buffer 540 µl 2.7 ml 8.1 ml 13.5 ml 27.0 ml

Vacuum Protocol
The vacuum pump shoold be a single
or double-staged unit capable of
producing up to 400 mm Hg pressure.
Centrifugation Protocol
A swinging bucket rotor is required for
  1. Ensure the connections of the Zymo-Spin™ III-S Column Assembly are finger-tight and place onto a vacuum manifold.
  2. Transfer the entire mixture from step 4 into the Zymo-Spin™ III-S Column Assembly. Switch on the vacuum pump until all the mixture has been completely drawn through the column.
  1. Ensure the connection between the Reservoir and Zymo-Spin™ III-S Column is finger-tight and place the assembly into a 50 ml conical tube.
  2. Transfer up to 10 ml of the mixture from step 4 into the Zymo-Spin™ III-S Column Assembly and centrifuge at 1,000 x g for 2 minutes. Discard the flow-through and repeat until the entire mixture has passed through the column.

Note: Steps 7-9 can also be completed using vacuum manifold instead of the microcentrifuge.

  1. Unscrew the orange Luer Lock cap from the top of the Zymo-Spin™ III-S Column and discard the top Reservoir.
  2. Place the Zymo-Spin™ III-S Column in a Collection Tube. Add 400 µl S&P DNA Prep Buffer to the Zymo-Spin™ III-S Column and centrifuge at ≥ 10,000 x g for 30 seconds in a microcentrifuge. Discard the flow-through.
  3. Add 700 µl S&P DNA Wash Buffer to the Zymo-Spin™ III-S Column and centrifuge at ≥ 10,000 x g for 30 seconds. Discard the flow-through.
  4. Add 400 µl S&P DNA Wash Buffer to the Zymo-Spin™ III-S Column and centrifuge at foll speed for 1 minute.
  5. Transfer the column into a 1.5 ml DNase-free tube and add ≥ 50 µl DNA Elution Buffer directly to the column matrix. Incubate for 3 minutes at room temperature and then centrifuge at maximum speed for 30 seconds.
For specific notes and additional information, please see the product protocol PDF.
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"Zymo Research’s Quick-cfDNA™ Serum & Plasma Kit was superior to Qiagen’s QIAmp Circulating NA Kit, customized both in fetal DNA recovery and in time savings.”
R. C. (DNA Diagnostics Center)
“… great yield of DNA!”
C. T. (University Hospital Carl Gustav Carus Fetscherstr)
“I liked the familiar layout, and the easy steps. I especially liked the reference tables that gave some sample volumes and calculations.”
A. U. (Cincinnati Children's Hospital)
“Higher DNA concentration in the final eluate from the Zymo kit when compared to the Qiagen kit. Total yield, measured by reference genes, is higher for the Zymo kit when compared to the Qiagen kit.”
T.R. (Aarhus University Hospital)
“Easy to use and good yield.”
X.Q (University of California, San Diego)
"Flexibil[e] for input volumes [with] quick references.”
E. L. (Cepheid)
Click here to submit your review of the Quick-cfDNA™ Serum & Plasma Kit.

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