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TruePrime® Apoptotic Cell-Free DNA Amplification Kit

The TruePrime® Apoptotic Cell-Free DNA Amplification Kit is optimized to exponentially amplify low input cell-free DNA (160-170bp) derived from apoptotic cells in liquid biopsy samples. One of the main challenges in cell-free DNA analysis is the limited amount of DNA obtained from bodily fluids, which affects the number and type of tests that can be performed. Further challenges are posed by short DNA fragment lengths, as well as the damaged nature of apoptotically-derived DNA. See how the TruePrime® Apoptotic Cell-Free DNA Amplification Kit provides solutions to these challenges.

 

 

Using multiple displacement amplification (MDA), accurate isothermal DNA amplification takes place, using cell-free DNA obtained from plasma, serum, urine, CSF or any other bodily fluids.

Note: This kit cannot be used for the amplification of DNA from necrotic cells (≥1kb). For this we would recommend the TruePrime® necrotic cell-free / exosomal  DNA amplification kit.

FeaturesBenefits
Integrated cell-free DNA pre-treatment (end-repair + dA-tailing reaction and ligation of hairpin-adaptors)Efficient amplification of apoptotic cell-free DNA without additional pre-treatment protocols and additional components
(cost and time saving)
Primer-free amplification – TthPrimPol synthesizes the primers for Phi29 DNA polymeraseNo primer-associated artefacts such as primer dimers – better representativity (cost and time saving)
Optimised for small template sizes (160-170 bp)Optimized kit and protocol for small templates (time saving)
Excellent sensitivity – down to picograms of input materialPerfect for precious / rare samples
Reliable at a cfDNA input level of 100 pg, preserving SNVs and SNV frequencies across different sequencing technologiesAmplification product sequencing compatibility
High amplification yieldsMore amplification product available for extended testing
(cost and time saving)
Excellent coverage with minimal biasReliable results for publication
Flexibility, up to 150 ng or 50 µl of cfDNA inputApplicational flexibility (time saving)
Wide variety of cell-free DNA sample sources
• Plasma
• Serum
• Urine
• CSF
• Other bodily fluids
Less restriction in sample source – greater experimental flexibility
Streamlined workflow and reduced hands-on timeCost and time saving

Applications:

  • Genetic research using small-fragmented (160-170bp) circulating cell-free DNA
  • Cancer research

Expected amplification yields

48 cancer patients were recruited and 10 mls of blood were extracted using Streck Cell-free DNA BCT® tubes. 3 mls of plasma were immediately isolated through a double-spin centrifugation protocol to avoid genomic DNA contamination from nucleated blood cells. Cell-free DNA was purified from 1ml of plasma using Qiagen QIAmp® Circulating Nucleic Acid Kit. Cell-free DNA was quantified using QubitTM. 1 ng of each cell-free DNA sample was subjected to the TruePrime® apoptotic cell-free DNA amplification workflow. The figure below shows the DNA amplification yields obtained in each case. All cell-free DNA samples were efficiently amplified, producing enough DNA for any subsequent analysis or technique.

TruePrime® apoptotic kit expected amplification yields

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