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EpiQuik Circulating Cell-Free DNA Isolation Kit

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For isolation of small fragment circulating cfDNA from plasma or serum

Citations (8) | (1) | Write a Review
Workflow of the EpiQuik™ Circulating Cell-Free DNA Isolation Kit.
High recovery of ccfDNA: Different amounts of HeLa mononucleosomes were spiked into 0.5 ml of plasma then isolated using the EpiQuik™ Circulating Cell-Free DNA Isolation Kit. The isolated DNA was fluorescently quantified using control DNA which was directly isolated from the same amount of mononucleosomes.
Input Type: Unisolated Samples
Research Area: DNA Damage & Repair, DNA Methylation, Next Gen Sequencing
Target Application: Sample Isolation
Vessel Format: Plate/Tubes
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-1064-2525 isolations $119.00 
P-1064-5050 isolations $205.00 
Order now & get it by Tuesday, August 27th  
Product Overview

We recommend a newer, more robust version of this kit: EpiQuik Circulating Cell-Free DNA Isolation Easy Kit

 

 

 

The EpiQuik™ Circulating  Cell-Free DNA Isolation Kit utilizes magnetic beads based size-fractionation technology to isolate circulating cell-free DNA (ccfDNA) from mono-nucleosomal and di-nucleosomal complexes in plasma/serum samples. The isolated ccfDNA can be directly used for real time-PCR and DNA library preparation suitable for next generation sequencing. The kit has the following advantages:

  • Uses innovative magnetic bead based size-fractionation technology for selective isolation of circulating cell-free DNA from plasma/serum that is mainly 170 bps in size. The isolated DNA can be directly used for both qPCR and NGS DNA library preparation.
  • Fast and straightforward procedure can be finished within 2 hours. No gels, columns or centrifugation is needed. 
  • Efficient removal of proteins, salts, nucleases, PCR inhibiting substances, and other impurities such as polysaccharides, polyphenols and lipids.
  • Sensitive and efficient DNA capture enables successful isolation with high recovery (>80% of input monoucleosomal DNA), even when the quantities of starting material are limited (as low as 0.1 ml). 
  • Manual and automation friendly – Scalable for single tube or 96-well plate formats.

Background Information
Genetic and epigenetic analysis of circulating cell-free DNA (ccfDNA) in plasma/serum or other body fluids provides unique opportunities for early detection of a wide range of clinical disorders such as cancer, autoimmune disease, infection and fetal disorders. It was demonstrated that ccfDNA of clinical importance occurs predominantly as fragments of approximately 170 bases from mononucleosomes with a smaller proportion as fragments of 360 bases from di-nucleosomes [1,2 ]. Such nucleosomal complexes are released into blood circulation during apoptotic cell death and will be increased under various pathological circumstances such as inflammation, pulmonary embolism, autoimmune disease, and cancer [3,4]. It is also shown that using ccfDNA from such nucleosomal complexes for genetic or epigenetic analysis provides better and more accurate identification of physiological and pathological status [5]. There are several methods currently being used for ccfDNA isolation from plasma and serum. All of these methods are based on capture of DNA by silicone column binding or phenol-chloroform separation. The DNA isolated by these methods contains both ccfDNA and non-ccfDNA, which may affect the accuracy of downstream analysis. To address these problems, Epigentek offers the EpiQuik™ Circulating Cell-Free DNA Isolation Kit for ccfDNA isolation. 

Principle & Procedure
The EpiQuik™ Circulating Cell-Free DNA Isolation Kit contains all components which have been optimized for the simple and rapid isolation of small size nucleosomal DNA from plasma/serum. The mono- and di-nucleosomal complexes are efficiently captured via size-fractionation magnetic beads (ccfDNA Capture Beads) by applying the beads to a magnetic field (EpiMag™ HT (96-Well) Magnetic Separator or similar). The captured nucleosomal DNA is then enzymatically released, and purified using MQ Binding Beads by simply washing the beads. The purified ccfDNA is then eluted from the beads for immediate use or storage.

Starting Materials
Both fresh and frozen plasma/serum from various sources can be used. However, fresh plasma/serum will generally give higher DNA yields than frozen. Furthermore, frozen plasma/serum will lead to DNA loss of about 10% per year. The input volume of plasma/serum can be from 0.1-1 ml with the standard volume of 0.5 ml per sample. If serum sample is used, the serum should be prepared within 6 hours after blood draw, since lysis of peripheral blood lymphocytes may cause an artificial increase in the amount of DNA during serum separation.  

Fig 4. High selectivity of specifically isolating small size ccfDNA (mononucleosomal DNA): Panel A: 500 ng of unspiked control polynucleosome (up to 2000 bps); Panel B: The 500 ng of the same polynucleosome spiked in 0.5 ml plasma; Panel C: 500 ng of mononucleosome spiked in 0.5 ml plasma; Panel D: 500 ng of polynucleosome and 500 ng of mononucleosome, which were simultaneously spiked into 0.5 ml plasma.


References

1. Jahr S et al: Cancer Res. 2001, 61: 1659-1665
2. Suzuki N et al: Clin Chim Acta. 2008, 387: 55-58
3. Holdenrieder S et al: Crit Rev Clin Lab Sci. 2009; 46: 1-24 
4. Schwarzenbach H et al: Nat Rev Cancer. 2011; 11: 426–437 
5. Chan KCA et al: Clinical Chem.  2004, 50: 88-92

Product Components

ccfDNA Capture Beads
ccfDNA Capture Enhancer*
Capture Buffer
Digestion Solution
Proteinase K*
MQ Binding Beads
Elution Buffer
User Guide

* Spin the solution down to the bottom prior to use.

 

User Guide & MSDS

[User Guide]*
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

[Safety Data Sheet]
Customer Reviews

Rating by s***@ciwemb.edu Verified Purchase Reviewed on: Friday 13 November, 2015
Application Description
We plan to identify colon cancer specific genetic mutations from patient plasma samples. DNA isolation was needed to prepare whole genome sequencing. This product was specific for DNA isolation from plasma and was used because of the low cost. Circulating plasma DNA was successfully isolated and enriched between 200-1000bp.

The Good: Isolated plasma DNA can be directly used for next generation sequencing

The Bad: Takes more time for handling the whole process

Procedural Details
Starting Material: Plasma

Follow exactly the kit instructions and final DNA elution volume is 20 ul.

Other Thoughts
Take care not to over dry the capture beads during the final elution step.

Review Source: http://www.biocompare.com/Product-Reviews/177177-Fast-and-excellent-Plasma-DNA-isolation/



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