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작성일 : 16-11-02
[HansaBioMed Life Sciences] Exo-DNAc, genomic DNA extraction kit from exosomes

 

Exo-DNAc, genomic DNA extraction kits from exosomes

Allows to concentrate the yield and to increase the purity of the DNA

 

 

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Circulating DNA is emerging as a novel non-invasive tool for patient's stratification and disease monitoring.

While most of the research has focused on circulating cell-free (cfDNA) or circulating-tumor-cell-(CTC)-

derived DNA, extracellular vesicle-(EVs)-associated DNA (EV-DNA) is emerging as a third valuable "liquid

biopsy" platform. In fact, genomic single or double-stranded DNA and mitochondrialDNA have been recently

detected in extracellular vesicles. In particular, the majorityof double-stranded DNA seems to be associated

with tumor derived exosomes (Thakur BK et al. 2014; Kalhert et al. 2014) where it represents the entire

genome of the cancerous tumor from which exosomes were derived. This discovery corroborates the

potential of exosomes, which can be easily obtained from a simple blood test.

 

The new EXO-DNAc Kit combines the ability of our reagent DNA-Prep to co-isolate EVs and circulating DNA

from biofluids (plasma, urine & serum, etc.) or culture supernatants with a user-friendly system for DNA

purification. Isolated vesicles are lysed with the appropriate lysis buffer and DNA is purified by spin columns

and optimized buffers with a fast turnaround time (approximatively 30 minutes). Finally,EXO-DNAc provides

an appropriated DNA concentrator for concentrating the yield (4folds) & increasing the purity of the DNA to the

high levels required for digital PCR analysis and NGS.

 

 

Features

• Highly pure circulating and EVs-associated DNA

• Fast and easy protocol

• Small volume amount required

• Suitable for concomitant profiling of cell-free circulating DNA and EV-associated DNA

 

 

Applications

• Discovering of mutations by qPCR and digital PCR

• Direct exosome capture and DNA purification from biofluids of cell media without time consuming

  purification steps.

• Isolation and profiling of genomic EV-associated DNA by DNAse treatment.

• DNA suitable for NGS

 

 

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Analysis can be limited to EV-associated DNA treating the pellet containing circulating DNA and EVs wit

Dnase. Extracellular vesicles protect the internal DNA from the Dnase digestion.

 

 

Exosome-associate DNA is suitable for point mutation analysis by allele-specific PCR

Healthy donor serum was spiked with 100 μg of purified exosome from BRAFV600E-positive A375 melanoma

cell lines. Vesicles and circulating DNA were isolated by precipitation with DNA-Prep contained in EXO-DNAc

kit and treated with or without Dnase 1, to distinguish circulating + Exosome related and only Exosome

related DNA. Following digestion, DNA was extracted with EXO-DNAc kit and analyzed by bioanalyisisand

allele-specific qPCR (Fig 1 and 2).

 

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Fig. 1.  Electropherograms of genomic DNA extracted with EXO-DNAc Kit with or without Dnase treatment

 

 

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Fig. 2. BRAF WT (wild type) and BRAF V600E amplification by qPCR from DNA extracted with EXO-DNAc Kit

with or without Dnase treatment.

 

 

EXO-DNAc is a flexible platform for detecting actionable mutations in cancer patients by

dPCR

Detection of the mutation BRAFV600E by digital PCR in genomic DNA isolated from serum of a metastatic

prostate cancer patient treated with abiraterone. Circulating and EVs associated DNA was extracted by EXO-

DNAc kit and a competitor kit. Digital PCR analysis revealed mutations in DNA extracted with EXO-DNAc,

whereas no mutation were detectable in DNA extracted with competitor kit. (Validated by Exosomics Siena)

 

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Ordering informations

    

Catalog No.

Description

Size

HBM-DNAc-PS-20/#

Isolation of circulating and exosome associated DNAc

from plasma and serum

20 reactions

HBM-DNAc-PS-40/#

40 reactions

HBM-DNA-PS-20/#

Isolation of circulating and exosome associated DNA

from plasma and serum (Old version)

20 reactions

HBM-DNA-PS-40/#

40 reactions

HBM-DNAc-UC-20/#

Isolation of circulating and exosome associated DNAc

from urine and cell culture media

20 reactions

HBM-DNAc-UC-40/#

40 reactions

HBM-DNA-UC-20/#

Isolation of circulating and exosome associated DNA

from urine and cell culture media (Old version)

20 reactions

HBM-DNA-UC-40/#

40 reactions

 

 

Exo-DNAc brochure (PDF)

HansaBioMed 2016 catalog (PDF)

 

 

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