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.

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.

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.

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.

Reversible terminator sequencing (Illumina) Whilst many other techniques use emulsion PCR to amplify the DNA library fragments, reversible termination uses bridge PCR, improving the efficiency of this stage of the process.

Pyrosequencing is used to reveal the genetic code of a section of DNA. It is also able to detect single nucleotide polymorphisms, insertion-deletions or other sequence variations, in addition to being able to quantify DNA methylation and allele frequency.

In genetics, shotgun sequencing is a method used for sequencing random DNA strands. It is named by analogy with the rapidly expanding, quasi-random shot grouping of a shotgun. Computer programs then use the overlapping ends of different reads to assemble them into a continuous sequence. …

PCR techniques play an integral role in targeted NGS sequencing, allowing for the generation of multiple NGS libraries and the sequencing of multiple targeted regions simultaneously.

Pyrosequencing is a method of DNA sequencing that detects light emitted during the sequential addition of nucleotides during the synthesis of a complementary strand of DNA.

DNA polymerase incorporates the correct, complementary dNTPs onto the template. This incorporation releases pyrophosphate (PPi). ATP sulfurylase converts PPi to ATP in the presence of adenosine 5´ phosphosulfate.

DOI: 10.1007/978-1-4939-3360-0_4 Abstract 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.

In contrast to ordinary emulsions, microemulsions form upon simple mixing of the components and do not require the high shear conditions generally used in the formation of ordinary emulsions. The three basic types of microemulsions are direct (oil dispersed in water, o/w), reversed (water dispersed in oil, w/o) and bicontinuous.

Microemulsions also have industrial applications, one of them being the synthesis of polymers. Microemulsion polymerization is a complex heterogeneous process where transport of monomers, free radicals and other species (such as chain transfer agent, co-surfactant and inhibitors) between the aqueous and organic phases, takes place.

Polymerization rate is controlled by monomer partitioning between the phases, particle nucleation, and adsorption and desorption of radicals. Particle stability is affected by the amount and type of surfactant and pH of dispersing medium. It is also used in the process of creating nanoparticles.