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BIO-0013 (103)
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20.2, 20.3 Class Notes.docx

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Department
Biology
Course
BIO-0013
Professor
Michelle Gaudette
Semester
Fall

Description
October 11, 2013 Needed for DNA: • Helicase • Primase • DNA polymerase III • dNTPs • topoisomerase • SSB proteins • DNA ligase • Telomerase (eukaryotes) • Clamp • DNA polymerase I • DNA template o Highlighted items are the ones absolutely necessary for DNA synthesis PCR is used to make millions of copies of a short DNA sequence • Synthesize primers in the laboratory so that you don’t need to synthesize primers o NEED to know the sequence of the DNA • How to design primers for PCR (DNA primers): o PCR primers must be located on either side of the target sequence, on opposite  strands  Make sure it’s read in the 5’  ▯3’ direction o When target DNA is single stranded, primers bind and allow DNA polymerase to  work  DNA can be made single­stranded by DNA helicases, or by heating to  temperatures that disrupt the hydrogen bonds • The logic behind PCR: o Denaturation of DNA duplex (94degreesC) o Annealing of primers (55­60degreesC) o DNA synthesis by 
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