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The DNA attaches to the flow cell via complementary sequences. The strand bends over and attaches to a second oligo forming a bridge. A polymerase synthesizes the reverse strand. The two strands release and straighten. Each forms a new bridge (bridge amplification). The result is a cluster of DNA forward and reverse strand clones.
Clonal Bridge Amplification Reversible Dye Terminator 2x300 0.17-2.7 15 Illumina HiSeq Clonal Bridge Amplification Reversible Dye Terminator 2x150 0.3-11 [12] 1000 [13] Illumina Genome Analyzer IIX Clonal Bridge Amplification Reversible Dye Terminator [14] [15] 2x150 2-14 95 Life Technologies SOLiD4 Clonal-emPCR Oligonucleotide 8-mer Chained ...
The technology features bridge amplification to generate clusters and reversible terminators for sequence determination. [52] [53] The technology behind these sequencing systems involves ligation of fragmented DNA to a chip, followed by primer addition and sequential fluorescent dNTP incorporation and detection.
However, other earlier patented technologies, such as that from Manteia Predictive Medicine (acquired by Solexa), which generate DNA on a solid-phase surface by bridge amplification, are generally referred to as "clusters". The terminology and distinction between 'polony' and 'cluster' have become confused recently.
Polony sequencing is a development of the polony technology from the late 1990s and 2000s. [5] Methods were developed in 2003 to sequence in situ polonies using single-base extension which could achieve 5-6 bases reads. [6] By 2005, these early attempts had been overhauled to develop the existing polony sequencing technology. [7]
[1] [2] Adams has made a notable contribution to the field of genetics as a co-inventor (with Steve Kron) of "bridge amplification," a polymerase chain reaction (PCR) technique that paved the way for development of DNA sequencing and genome sequencing. [3]
The sequencing methods applied by these sequencers do not require DNA amplification (polymerase chain reaction – PCR), which speeds up the sample preparation before sequencing and reduces errors. In addition, sequencing data is collected from the reactions caused by the addition of nucleotides in the complementary strand in real time.
Multiple Annealing and Looping Based Amplification Cycles (MALBAC) is a quasilinear whole genome amplification method. Unlike conventional DNA amplification methods that are non-linear or exponential (in each cycle, DNA copied can serve as template for subsequent cycles), MALBAC utilizes special primers that allow amplicons to have complementary ends and therefore to loop, preventing DNA from ...