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.
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.
| Quick-cfDNA™ Serum & Plasma Kit |
50 preps |
D4076 |
 |
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