DNA sequencing determining the order of nucleotides in

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DNA sequencing determining the order of nucleotides in DNA – Sanger (di-deoxy) 600 -1000

DNA sequencing determining the order of nucleotides in DNA – Sanger (di-deoxy) 600 -1000 bp – Next-Generation Sequencing • 2 nd generation • 3 rd generation 454 -Pyrosequencing • Solexa/Illumina Secuencing • True Single Molecule Sequencing • Ion Torrent

Solexa/Illumina Secuencing The sequencing-by-synthesis process.

Solexa/Illumina Secuencing The sequencing-by-synthesis process.

True Single Molecule Sequencing • • • By directly sequencing single molecules of DNA

True Single Molecule Sequencing • • • By directly sequencing single molecules of DNA or RNA, Helicos' True Single Molecule Sequencing (t. SMS™) and Direct RNA Sequencing (DRS™) technologies significantly increase the speed of sequencing, while also decreasing the cost. Polymerase and one fluorescently labeled nucleotide (C, G, A or T) are added. billions of templates

Single Molecule Real Time Sequencing • Single molecule real time sequencing utilizes the zero-mode

Single Molecule Real Time Sequencing • Single molecule real time sequencing utilizes the zero-mode waveguide (ZMW) • A single DNA polymerase enzyme is affixed at the bottom of a ZMW with a single molecule of DNA as a template • Each of the four DNA bases is attached to one of four different fluorescent dyes. • SMRT chip contains ~3000 ZMW holes • Each of the ZMW holes produces approximately 1, 500 bp (base pair) read lengths at a speed of 10 bp per second

Single Molecule Real Time Sequencing

Single Molecule Real Time Sequencing

Ion Torrent • Fastest sequencing workflow • Two hour sequencing run for up to

Ion Torrent • Fastest sequencing workflow • Two hour sequencing run for up to 200 bp reads • Fully prep 8 samples in parallel in less than 6 hours

The Illumina genotyping platform

The Illumina genotyping platform

Nano. Drop • The Nano. Drop® ND-1000 is a fullspectrum (220 -750 nm) spectrophotometer

Nano. Drop • The Nano. Drop® ND-1000 is a fullspectrum (220 -750 nm) spectrophotometer that measures 1 ul samples with high accuracy and reproducibility. • Nucleic acid concentration and purity of nucleic acid samples up to 3700 ng/ul (ds. DNA) without dilution • Fluorescent dye labeling density of nucleic acid microarray samples • General UV-Vis spectrophotometry

Nano. Drop • 1 ul only of sample required * Large dynamic range: 23700

Nano. Drop • 1 ul only of sample required * Large dynamic range: 23700 ng/ul of ds. DNA (manufacturer's specs, this is probably pushing it at both ends) * No dilutions for most samples * No cuvettes or capillaries * 10 second measurement • DNA, RNA, microarray labeling dyes, and proteins

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