These ddntps will also be radioacnvely for detecnon

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Unformatted text preview: ors SubsNtuNon 1 Billion Illumina HISeq 150 Million 100 – 200bp 15 Billion SubsNtuNon with exponenNal increase PacBio 1000 ­2000bp 45 Million InserNons and deleNons 45,000 \ Sequencing Methods and Terminology Sanger Sequencing •  The key principle of the Sanger method was the dideoxynucleoNde triphosphates (ddNTPs) as DNA chain terminators. •  These ddNTPs will also be radioacNvely for detecNon in automated sequencing machines. •  PosiNves: longer reads (600 to 1000 bp). •  NegaNves: poor coverage (6x), expensive, inaccurate. •  SNll commonly used for small scale sequencing. Sanger Sequencing Video Sanger Sequencing DNA target sample SHEAR Sanger Sequencing DNA target sample A T C G SHEAR A T C G Close each fragment many times. A T C G A T C G Sanger Sequencing DNA target sample SHEAR T A T A C G A T C C G A T C G A T C G G 28 Sanger Sequencing DNA polymerase A T A C G Primer Sanger Sequencing DNA polymerase A T A C G Primer Primer A T A C G DNA polymerase Sanger Sequencing A G C Primer DNA polymerase C G A C T A C T A C T A T A C G Sanger Sequencing A G C Primer G DNA polymerase C G A C T A C T A C T A T A C G Sanger Sequencing A G C C G A C T A C T A C T Primer G G A T A C G Sanger Sequencing A G C C G A C T A C T A C T Primer G G C A T A C G Sanger Sequencing A G C C G A C T A C T A C T Primer G G C T G A A T A C G Sanger Sequencing A G C C G A C T A C T A C T Primer S...
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This note was uploaded on 02/10/2014 for the course CS 680 taught by Professor Staff during the Fall '08 term at Colorado State.

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