Changes
From Biolecture.org
no edit summary
<p><span style="font-size: medium"><b>Synthesis based sequencing by Illumina</b></span></p>
<p><font size="3">Solexa, now part of Illumina developed a sequencing technology based on reversible dye-terminators. DNA molecules are first attached to primers on a slide and amplified so that local clonal colonies are formed (bridge amplification). Four types of ddNTPs are added, and non-incorporated nucleotides are washed away. Unlike pyrosequencing, the DNA can only be extended one nucleotide at a time. A camera takes images of the fluorescently labeled nucleotides then the dye along with the terminal 3' blocker is chemically removed from the DNA, allowing a next cycle. The final product of the SBS is many [[DNA reads]].</font> </p>
<p> </p>
<p> </p>
<p><b style="font-size: medium;">ligation based sequencing </b></p>
<p> </p>
<p><span style="font-size: medium;">In this method, octomer-oligonucleotide probe is used, and instead of DNA polymerase, DNA ligase is used, as its name implies. Each of probes are dyed according to its first two nucleotides. First, primer is annealed to the template strand, probe follows, and DNA ligase is applied on the junction. unbound probe is washed away, and signal from bound probe is detected. End of probe is cleaved away from 3 nucleotide behind from first two nucleotides. Cycle is repeated with another primer, which has different starting base. The data are collected and compared to compensate empty nucleotides. This method is usually used for short strands. </span></p>
<p> </p>
<p> </p>
<h2><span class="mw-headline">RNA sequencing</span></h2>
