Lecture_31 Sequencing

Lecture_31 Sequencing - ANNOUNCEMENTS Apr 9 Stop by Janes...

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ANNOUNCEMENTS – Apr. 9 • Next reading quiz covers Chapter 21 , due before class Mon., Apr. 14. Make-up Midterm Exam 2 , Friday, April 11, 5:30-7:00, 106 BBE Must have preapproval of an instructor. Volunteers needed! Get your own clicker out and ready. Loaners available if you need one. Stop by Jane’s Addiction
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Lect. 31 Sequencing & Genomics
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Sequencing & Genomics: Learning Goals Sequencing a DNA template is accomplished by assessing the incorporation of individual bases PCR or cloning is used to generate large quantities of identical template DNA for sequencing
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Sequencing & Genomics: Key Terms
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DNA Sequencing & Genomics: Learning Goals & Objectives Sequencing a DNA template is accomplished by assessing the incorporation of individual bases § Interpret the base sequence and polarity from products of a sequencing reaction on a single template § Recognize the components and processes used in Sanger sequencing using dye terminators
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17.1 How is DNA Sequenced? Current methods of DNA sequencing rely on measuring the sequential incorporation of bases as DNA is synthesized. One common method uses chemically modified nucleosides (ddNTPs) in combination with normal nucleosides (dNTPs). Synthesis stops when a ddNTP is added by DNA polymerase to a polynucleotide chain because ddNTP has no hydroxyl group at the 3 position. The sequence is determined by detecting the terminal (3 ) base (A, C, G, or T) at each position in the DNA.
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Figure 17.2 Sequencing DNA synthesized strand template strand
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17.1 How Is DNA Sequenced? Components of sequencing reactions: q DNA polymerase q Short (18-22 bases) primer complementary to template strand q All 2 deoxynucleoside triphosphates (dATP, dCTP, dGTP, dTTP) q All 2 ,3 dideoxynucleoside triphosphates (ddATP, ddCTP, ddGTP, ddTTP); labeled uniquely so that each can be detected q Template DNA, sequencing reaction reliably read only 100-700 bases of a template
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17.1
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