Indirect Sequence Capture

Capture long DNA targets >100 kb.
No target amplification.
No target hybridization.

Indirect Sequence Capture – what it is and what it’s not

1

ENCAPSULATES single long DNA molecules in droplets with primers for the Detection Sequence.

2

LABELS TARGETS by amplifying a ~150 bp Detection Sequence within or flanking the target.

3

SORTS out droplets containing the target based on the fluorescent signal of the amplified Detection Sequence.

indirect sequence capture ISC

Indirect sequence capture (Xdrop™)


DECOUPLES ENRICHMENT FROM AMPLIFICATION OR HYBRIDIZATION

  • no need to know the target sequence
  • avoids PCR bias
  • works with low sample input

indirect sequence capture aplicon based

Amplicon based

indirect sequence capture Hybridization capture

Hybridization-capture

Where you put the primers opens all kinds of possibilities

WITHIN THE TARGET

  • verification of insertions, gene duplications, and conserved regions where expected and not!
  • gene/pseudogene discrimination
  • haplotype phasing
  • CRISPR editing assessment

Known insertion

slides for tech page target knowninsertion

Unknown insertion

slides for tech page target unknowninsertion

Target gene

slides for tech page target target gene

Pseudogene

slides for tech page target pseudogene

FLANK THE TARGET

  • gap closing
  • resolving tandem repeats

Known region 1

???

Known region 2

slides for tech page target knownregion

Tandem repeat

slides for tech page target tandem repeat

Here’s how it works in detail

how it works step01

Xdrop workflow arrow ned

1

Mix High Molecular Weight (HMW) DNA, Primers and droplet PCR Reagents

Primers used for Target detection, not for Target amplification

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how it works icon syringe

how it works step02

Xdrop workflow arrow ned

2

Encapsulate DNA

Use Xdrop™ to encapsulate DNA fragments into millions of droplets


how it works icon machine

how it works step03

Xdrop workflow arrow ned

3

Detection Sequence Amplification

A short (~150 bp) sequence that identifies the Target DNA is amplified


how it works icon machine2

how it works step04

Xdrop workflow arrow ned

4

Stain

Target-positive droplets are stained green


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how it works step05

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5

Select Target-positive Droplets

A cell sorter is used to isolate green-stained positive droplets


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how it works step06

6

Release HMW Target DNA

Break solution is added to release Target native DNA from droplets

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how it works icon syringe


Brochure workflow SAMPLIX A3

See how droplet PCR works

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