* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download here
Human genome wikipedia , lookup
DNA paternity testing wikipedia , lookup
Designer baby wikipedia , lookup
DNA methylation wikipedia , lookup
DNA barcoding wikipedia , lookup
Nutriepigenomics wikipedia , lookup
Zinc finger nuclease wikipedia , lookup
Site-specific recombinase technology wikipedia , lookup
DNA sequencing wikipedia , lookup
Comparative genomic hybridization wikipedia , lookup
Holliday junction wikipedia , lookup
Mitochondrial DNA wikipedia , lookup
DNA profiling wikipedia , lookup
Point mutation wikipedia , lookup
Genomic library wikipedia , lookup
Cancer epigenetics wikipedia , lookup
Microevolution wikipedia , lookup
No-SCAR (Scarless Cas9 Assisted Recombineering) Genome Editing wikipedia , lookup
Vectors in gene therapy wikipedia , lookup
SNP genotyping wikipedia , lookup
DNA vaccination wikipedia , lookup
DNA nanotechnology wikipedia , lookup
DNA damage theory of aging wikipedia , lookup
Genealogical DNA test wikipedia , lookup
Gel electrophoresis of nucleic acids wikipedia , lookup
Bisulfite sequencing wikipedia , lookup
United Kingdom National DNA Database wikipedia , lookup
Non-coding DNA wikipedia , lookup
Microsatellite wikipedia , lookup
Molecular cloning wikipedia , lookup
Cell-free fetal DNA wikipedia , lookup
Epigenomics wikipedia , lookup
Primary transcript wikipedia , lookup
Therapeutic gene modulation wikipedia , lookup
DNA replication wikipedia , lookup
History of genetic engineering wikipedia , lookup
Artificial gene synthesis wikipedia , lookup
Extrachromosomal DNA wikipedia , lookup
Nucleic acid double helix wikipedia , lookup
DNA polymerase wikipedia , lookup
Nucleic acid analogue wikipedia , lookup
DNA supercoil wikipedia , lookup
Cre-Lox recombination wikipedia , lookup
Helitron (biology) wikipedia , lookup
Today’s Outline • Study Notes 6b Due! • Take up exam – View exam on Friday between 2-4pm in C-4. – Written appeals-process • DNA Replication – Replication – PCR – Telomeres • DNA Replication Tutorial 3-5pm Friday Write Down: What is one thing you learned about DNA replication from the readings? To Understand DNA Replication, Understand the Parts - Helicase Topoisomerase Single Strand Binding Proteins DNA Polymerase III Sliding Clamp DNA Primase Helicase Helicase unzips DNA. Topoisomerase Relieves the tension produced when DNA is unzipped. Single Strand Binding Proteins Stabilizes DNA and prevents it from binding to parallel strand. DNA Polymerase III Adds dNTPs to the newly synthesized strand on the leading strand. Adds in a 5’ 3’ direction. Sliding Clamp Holds DNA Polymerase III in place. DNA Primase Nucleotides can ONLY be added to the 3’ end of an already-attached nucleotide. A primer provides this. DNA Primase builds the primer. DNA Polymerase Chain Reaction • What does polymerase do? • What if…. – I put an entire genome in a test tube. – I added DNA polymerase III and dNTPs. – I added very specific DNA primers that mapped onto a specific gene. – Then I started heating and cooling the DNA over and over and over and over…. Polymerase Chain Reaction 5’ 3’ 3’ 5’ 5’ 3’ 3’ 5’ 5’ 3’ 5’ 3’ - taq polymerase dNTPs primers buffer Discuss: • Find three similarities and three differences between (i) DNA replication on the leading strand and (ii) DNA replication via PCR. (Debrief using name cards) The 1st Challenge of Lagging Strand Replication • DNA Primase is used over and over • DNA Primase lays down ribonucleic acid. Solution: an enzyme that can replace RNA nucleotides with DNA nucleotides DNA Pol1 The 2nd Challenge of Lagging Strand Replication Solution: an enzyme that can connect 3’ carbons to phosphate groups LIGASE 5 Ends of parental DNA strands Leading strand Lagging strand 3 3 5 Last fragment Previous fragment Lagging strand RNA primer 5 3 Parental strand Removal of primers and replacement with DNA where a 3 end is available DNA Pol1 Can only add nucleotides to the 3’ end of existing DNA. 5 3 Telomeres protect an organism’s genes from being eroded during successive rounds of DNA replication. In humans the sequence TTAGGG is repeated between 100 and 1000 times. Second round of replication 5 New leading strand 3 New lagging strand 5 3 Further rounds of replication Shorter and shorter daughter molecules Which of these replaces RNA with DNA? A) B) C) D) E) DNA Polymerase 1 DNA Polymerase 3 DNA Primase Ligase Nuclease Which of these glues 3’ and 5’ DNA ends together? A) B) C) D) E) Ligase DNA Polymerase 1 Binding Proteins DNA Polymerase 3 DNA Primase Which of these adds new nucleotides to the leading strand? A) B) C) D) E) DNA Polymerase 3 The Sliding Clamp DNA Polymerase 1 DNA Primase Binding Proteins Which of these is not needed by the leading strand: A) B) C) D) E) DNA Polymerase 1 Binding Proteins DNA Primase The Sliding Clamp DNA Polymerase 3 Which of these is not needed by the lagging strand: A) B) C) D) E) The Sliding Clamp DNA Polymerase 1 Binding Proteins DNA Polymerase 3 All are needed Why are telomeres needed? a) Primers can’t bind to 5’ ends of DNA b) DNA polymerase 1 can’t add nucleotides to 5’ ends of DNA c) Okazaki fragments don’t form at the end of DNA strands d) Helicase can’t bind to the end of DNA strands. e) None of the above. Write Down: 1. On a scale of 1 to 10, how well do you understand DNA replication? 2. What is one thing you understand really well? 3. What is one thing you do not understand? Coming Up: 1. 2. 3. 4. 5. Sanger Sequencing (Tues) Molecular Sculpting of DNA replication (Tues) Bonus assignment due (see Lecture Schedule) Field trip sign up DNA Replication Tutorial 3-5pm Friday