What is emulsion PCR?

Emulsion PCR (ePCR) is an important technique that permits amplification of DNA molecules in physically separated picoliter-volume water-in-oil droplets, and thus avoids formation of unproductive chimeras and other artifacts between similar DNA sequences.

How was emulsion PCR used during genome sequencing?

This emulsion PCR (ePCR) method allowed spatial isolation of each template to a single bubble and enabled clonal amplification of the template on each bead. In 2005, two ground-breaking studies built on these discoveries and described high-throughput methods for rapidly and cheaply sequencing a whole bacterial genome.

What is EM PCR?

Emulsion PCR (EmPCR) is a commonly employed method for template amplification in multiple NGS-based sequencing platforms. The basic principle of emPCR is dilution and compartmentalization of template molecules in water droplets in a water-in-oil emulsion.

Which method is used to generate clonally amplified DNA fragments to be used as templates in Pyrosequencing?

emulsion PCR
DNA fragments are first attached to microbeads and clonally amplified in an emulsion PCR. The beads are consequently spread into individual picolitre-sized wells and subjected to pyrosequencing.

What is bridge PCR sequencing?

Bridging PCR (BPCR) is a combination of two processes, a recombination between two template sequences and an amplification of the recombinant template. Two parental sequences share a homologous region (a region in which the sequence is identical) and diverge in the nonhomologous flanking sequences.

What is bridge amplification in Illumina sequencing?

Bridge amplification takes place in a flow cell, aiming to generating clusters of DNA strands for further sequencing and analysis. By repeating this denaturation and extension process, millions of fragments are amplified, forming localized clusters on the flow cell.

How does SMRT sequencing work?

At the heart of SMRT Sequencing is the SMRT Cell, which contains millions of tiny wells called zero-mode waveguides (ZMWs). Single molecules of DNA are immobilized in these wells, and as the polymerase incorporates each nucleotide, light is emitted, and nucleotide incorporation is measured in real time.

What is nanopore sequencing technology?

Nanopore sequencing is a unique, scalable technology that enables direct, real-time analysis of long DNA or RNA fragments. It works by monitoring changes to an electrical current as nucleic acids are passed through a protein nanopore. The resulting signal is decoded to provide the specific DNA or RNA sequence.