Biochemistry - Bonham Chemistrybonhamchemistry.com/wp-content/uploads/2017/01/Lecture_3.pdfPrimer Template of unknown sequence DNA polymerase, four dNTPs, four ddNTPs Denature Dye-labeled

Biochemistry

Lecture 3 DNA Technologies

DNA  Cloning  

3  

Introduc2on    into  

host  cell  

Restriction Enzymes

PCR Polymerase Chain Reaction

Antibiotic Selection •  Antibiotics, such as penicillin and ampicillin, kill

bacteria •  Plasmids can carry genes that give host

bacterium a resistance against antibiotics •  Allows growth (selection) of bacteria that have

taken up the plasmid

NH

S

N

N

OO

H

OOH

HH

Expression of Cloned Genes

DNA Electrophoresis

Sanger  DNA  Sequencing  

14  

DNA Sequencing

Automated  DNA  Sequencing  

16  

Shotgun Sequencing

Applica2ons  of  Sequencing  

•  Solving  crimes  •  Iden2fying  gene2c  diseases  – Hun2ngton’s  disease  –  Breast  cancer  –  Sickle  cell  anemia  

•  Viral  infec2ons  •  Human  migra2on  paFerns  

•  Parentage/pedigree/genealogy  

18  

Genome  Edi2ng:  CRISPR-­‐Cas9  System  

19  Doudna,  JA,  Charpen2er  E.  Science.  2014;  346(6213).  

Electrochemical Sequencing

http://www.youtube.com/watch?v=yVf2295JqUg