ENGR213_Lecture6_BRAE213

ENGR213_Lecture6_BRAE213 - Lecture 6 PCR A deeper Look PCR...

Info iconThis preview shows pages 1–10. Sign up to view the full content.

View Full Document Right Arrow Icon

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
This is the end of the preview. Sign up to access the rest of the document.

Unformatted text preview: Lecture 6 PCR A deeper Look PCR a quick Review DNA Melting and Hybridization • Typical Temperatures: – Melting ~90 - 95 C – Hybridization ~40 - 60 C DNA The Genetic Code for Life Some new Information • The hydrogen bonds can be modified with heat, i.e, – DNA Melting Temperature – DNA Hybridization t m ≈ 64.9 + 0.41 % GC ( 29- 500 Length of DNA PCR a quick Review • Replication of a unique region of the genome to – Identify an organism – Forensics – Gene replication – Development of a cDNA library Temperature Cycle: ~95 C melting ~50 C Annealing Ramp ~70 C Extension PCR a quick Review First Cycle Subsequent Cycles: •Replication is a 2 n process where n is the number of cycles What else are we lacking? • Polymerase w and w/o Exonuclease • Primers (what’s that?) • Probes with “Reporters” • Reagents • Inhibitors New Definitions • Amplicon -- ssDNA that is amplified via DNA Polyermase • Exonuclease -- Enzyme that clips off any unpaired bases “proof reading” • Primer -- short, single strand of RNA that locates the start and end of a replication region for PCR • Probe -- piece of RNA or DNA labeled with Reporter that locates a specific region of DNA by hybridization Some New Definitions • Reporter -- A Radioactive or Fluorescent signal that is detectable • Exogenous DNA -- DNA outside an organism • Cycle -- One cycle of PCR replication • PCR-- Polymerase Chain Reaction • QPCR -- Quantitative PCR • RT-PCR -- Reverse Transcriptase PCR • real time-PCR -- Real Time PCR • ICC-PCR -- Integrated Cell Culture PCR Polymerases Polymerase Exo- nuclease General Info (Prokaryotic) Taq No Thermphilus Aquaticus; bacteria grows in Hot Springs Pfu Yes Pyrococcus furiosus, an Archea which thrives at 100  C, first...
View Full Document

This note was uploaded on 04/24/2008 for the course ENGR 213 taught by Professor Clague during the Winter '08 term at Cal Poly.

Page1 / 40

ENGR213_Lecture6_BRAE213 - Lecture 6 PCR A deeper Look PCR...

This preview shows document pages 1 - 10. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online