Fredrick Sanger <ul><li>developed dideoxy termination sequencing method </li></ul><ul><li>Nobel Laureate, 1980 </li></ul><...
DNA Sequencing <ul><li>similar to DNA replication </li></ul><ul><li>somehow stop at each base  </li></ul><ul><li>determine...
Sequencing Requirements <ul><li>DNA polymerase </li></ul><ul><li>DNA template strand </li></ul><ul><li>primer (radioactive...
deoxyribose when oxygen removed dideoxyribose when both oxygen atoms removed 3’ OH functional group necessary for DNA elon...
Sequencing Setup <ul><li>one test tube for each dideoxyribonucleotide  (ddATP, ddGTP, ddTTP, ddCTP) </li></ul>A G T C 15% ...
Sequencing Setup <ul><li>one test tube for each dideoxyribonucleotide  (ddATP, ddGTP, ddTTP, ddCTP) </li></ul>A G T C 15% ...
Sequencing Setup components in one test tube
After Replication Process <ul><li>contents of each tube is separated by gel electrophoresis </li></ul><ul><li>Look at your...
 
SANGER SEQUENCING http://www.dnalc.org/view/15479-Sanger-method-of-DNA-sequencing-3D-animation-with-narration.html Insulin...
Upcoming SlideShare
Loading in …5
×

17 sequencing-2010 update

562 views

Published on

Published in: Technology
0 Comments
0 Likes
Statistics
Notes
  • Be the first to comment

  • Be the first to like this

No Downloads
Views
Total views
562
On SlideShare
0
From Embeds
0
Number of Embeds
1
Actions
Shares
0
Downloads
0
Comments
0
Likes
0
Embeds 0
No embeds

No notes for slide

17 sequencing-2010 update

  1. 2. Fredrick Sanger <ul><li>developed dideoxy termination sequencing method </li></ul><ul><li>Nobel Laureate, 1980 </li></ul><ul><li>Nobel Laureate, 1958; determined insulin sequence </li></ul>
  2. 3. DNA Sequencing <ul><li>similar to DNA replication </li></ul><ul><li>somehow stop at each base </li></ul><ul><li>determine which was the last base added </li></ul>
  3. 4. Sequencing Requirements <ul><li>DNA polymerase </li></ul><ul><li>DNA template strand </li></ul><ul><li>primer (radioactive) </li></ul><ul><li>deoxyribonucleotides </li></ul><ul><li>dideoxyribonucleotides </li></ul>
  4. 5. deoxyribose when oxygen removed dideoxyribose when both oxygen atoms removed 3’ OH functional group necessary for DNA elongation
  5. 6. Sequencing Setup <ul><li>one test tube for each dideoxyribonucleotide (ddATP, ddGTP, ddTTP, ddCTP) </li></ul>A G T C 15% ATP 25% ATP 25% ATP 25% ATP 25% GTP 15% GTP 25% GTP 25% GTP 25% TTP 25% TTP 15% TTP 25% TTP 25% CTP 25% CTP 25% CTP 15% CTP 10% ddATP 10% ddGTP 10% ddTTP 10% ddCTP
  6. 7. Sequencing Setup <ul><li>one test tube for each dideoxyribonucleotide (ddATP, ddGTP, ddTTP, ddCTP) </li></ul>A G T C 15% ATP 25% ATP 25% ATP 25% ATP 25% GTP 15% GTP 25% GTP 25% GTP 25% TTP 25% TTP 15% TTP 25% TTP 25% CTP 25% CTP 25% CTP 15% CTP 10% ddATP 10% ddGTP 10% ddTTP 10% ddCTP
  7. 8. Sequencing Setup components in one test tube
  8. 9. After Replication Process <ul><li>contents of each tube is separated by gel electrophoresis </li></ul><ul><li>Look at your notes. </li></ul>
  9. 11. SANGER SEQUENCING http://www.dnalc.org/view/15479-Sanger-method-of-DNA-sequencing-3D-animation-with-narration.html Insulin http://www.dnalc.org/view/16515-Animation-23-A-gene-is-a-discrete-sequence-of-DNA-nucleotides-.html

×