droplet Multiple Displacement Amplification

Unbiased amplification of single long DNA sequences
containing the target.

Uniform sequence coverage.

No inter-template chimeras.

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The key is that dropMDA happens in droplets

mdma workflow

Partitioning one single DNA molecule in each droplet extremely reduces amplification bias:

  • no preferred amplification of certain templates
  • no nested products
  • no inter-template chimera

dMDAplot

Amplifying every template in separate droplets (dropMDA) provides a highly uniform sequence coverage, while bulk MDA preferentially amplifies certain templates.

And why MDA?

    • MDA amplifies any DNA, employing no specific primers (uses random hexamer primers)
    • The high-fidelity, proofreading Phi29 enzyme has a 10-fold lower error rate than Taq polymerase
    • The isothermal reaction leaves more template DNA intact
    • MDA preferentially amplifies sequences 1–70 kb long

mda cga

Hexamer primers bind at Multiple sites. Growing strands are Displaced by others, opening additional primer annealing sites to further boost Amplification.

Here’s how it works in detail

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7

Encapsulate Target DNA

Xdrop™ generates thousands of droplets, each with single Target DNA + amplification reagents


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8

Unbiased Samplicon Production

Each molecule of Target DNA undergoes multiple displacement amplification in droplets


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9

Release of ~ 1µg of Target DNA

Break solution releases Samplicons (amplified Target DNA) into solution

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Downstream Analysis

Target is enriched 100-1000x and is ready for long- and short-read sequencing, qPCR etc.

Ready for applications....

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See the whole flow

Brochure workflow SAMPLIX A3

See how droplet MDA works

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