cfDNA isolation from plasma

Automated isolation of circulating cell-free tumor or fetal DNA from 0.5 - 5 ml plasma

In the event of apoptosis, necrosis or break-down of pathogens, or cells, such as white blood cells circulating cell-free DNA (cfDNA) is floating through the blood. As the average size of cfDNA is 170 bp (50 – 200 bp) and only 1 – 50 ng of cfDNA/ml of plasma is present, it is quite difficult to separate this hay in the haystack. Nevertheless the analysis of cfDNA in blood is an elegant non-invasive method to study cancer or chromosomal disorders. PerkinElmer chemagen has a new method to isolate cfDNA form 0.5 - 5 ml plasma with minimal hands-on time with the chemagic instruments.

Product overview for automated cfDNA isolation

Catalog No.Kit NameInstrumentchemagic
Rod Head
FormatPreps/KitSample VolumesYieldProcessing Time
CMG-1444chemagic
Prime™ cfDNA 5k Kit H24
chemagic Prime 4/82424-well2403 - 5 ml1 - 30 ng/ml plasma (qPCR)240 min
CMG-1304chemagic
cfDNA 5k Kit H24
chemagic 360/MSM I2424-well2403 - 5 ml1 - 30 ng/ml plasma (qPCR)100 min
(incl. 15 min hands-on time)
CMG-1302chemagic
cfDNA 2k Kit H24
chemagic 360/MSM I2424-well2401 - 2 ml1 - 30 ng/ml plasma (qPCR)90 min
(incl. 15 min hands-on time)
CMG-1396chemagic
cfDNA 1.5k Kit H96
chemagic 360/MSM I9696-well9600.5 - 1.5 ml1 - 30 ng/ml plasma (qPCR)120 min
(incl. 30 min hands-on time)

Product overview for manual cfDNA isolation

Catalog No.Kit NameAccessoriesInputTypical YieldPreps/Kit
CMG-134chemagic™ cfDNA 5k Kitchemagic™ Stand 2 x 12
chemagic™ Stand 50k Type B
1 - 5 mldepending on the sample material40

Product Specifications

The kits are designed for the use with human plasma samples derived from EDTA, citrate or Streck® Cell-free DNA BCT® tubes. Fresh and frozen plasma can be used.

Technology

Technology

The chemagic™ cfDNA Kits are based on chemagic™ Technology using M-PVA Magnetic Beads for the isolation of cfDNA. The cfDNA binds to paramagnetic beads which are magnetically separated from the sample material. During subsequent steps contaminants are removed and the purified cfDNA is transferred into an elution medium. The automated sample processing by the chemagic™ 360 instrument or the chemagic Prime™ instrument excludes cross contamination and ensures safe handling of infectious sample material.

Plasma Preparation Protocol

Plasma Preparation Protocol

It is recommended to prepare plasma as fresh as possible (max. 5 days after blood draw). Longer storage of blood prior to plasma preparation may lead to poor separation results and higher background from genomic DNA.
A double centrifugation protocol during plasma preparation is recommended to minimize the potential for contamination of plasma with cells and genomic DNA. Transfer of any other blood components (buffy coat or red blood cells) should be avoided while separating the plasma fraction.
Please refer to the tube manufacturers specifications (max. centrifugation speed) for more information.
From 10 ml of whole blood approximately 4 - 5 ml plasma can be expected.

  1. Centrifuge whole blood collection tube at 2000 x g for 20 minutes.
  2. Aspirate plasma carefully and at least 2 - 3 mm above the buffy coat layer, without disturbing the layer and transfer it into a new appropriate tube.
  3. Centrifuge the plasma sample at 3300 x g for 30 minutes.
  4. Carefully transfer the supernatant to a fresh tube without disturbing the pellet, at least 2 - 3 mm above the pellet.
  5. For direct use storage of plasma sample at 2 - 8 °C is possible, for long-term storage -80 °C is recommended.

Download Manual

Catalog No.Kit NamePDF Protocol
CMG-134chemagic™ cfDNA 5k Kit (manual)Protocol CMG-134 chemagic cfDNA 5k Kit
CMG-1304chemagic™ cfDNA 5k Kit H24Protocol CMG-1304 chemagic cfDNA 5k Kit H24
CMG-1302chemagic™ cfDNA 2k Kit H24Protocol CMG-1302 chemagic cfDNA 2k Kit H24
CMG-1396chemagic™ cfDNA 1.5k Kit H96Protocol CMG-1396 chemagic cfDNA 1.5k Kit H96

For research use only. Not for use in diagnostic procedures.