BME100 f2013:W1200 Group11 L4: Difference between revisions
Meera Doshi (talk | contribs) |
Meera Doshi (talk | contribs) |
||
Line 148: | Line 148: | ||
''''''Diagram of PCR''''''<br> | ''''''Diagram of PCR''''''<br> | ||
[[Image:121312.jpg]]<br> There are two primers, the forward and the reverse primer, which bind to the DNA template. This image illustrates the reverse primer binding to the template. The colors of the circular components which bind to each other represent the different types of nucleotide bases DNA: adenine, guanine, cytosine, and thymine. The adenine base in the primer binds to the thymine base in the DNA template, the guanine base in the primer binds to the cytosine base in the DNA template. Likewise, the thymine in the primer binds to the adenine in the template, and the cytosine in the primer binds to the guanine in the template.<br> | [[Image:121312.jpg]]<br> There are two primers, the forward and the reverse primer, which bind to the DNA template. This image illustrates the reverse primer binding to the template. The colors of the circular components which bind to each other represent the different types of nucleotide bases in DNA: adenine, guanine, cytosine, and thymine. The adenine base in the primer binds to the thymine base in the DNA template, the guanine base in the primer binds to the cytosine base in the DNA template. Likewise, the thymine in the primer binds to the adenine in the template, and the cytosine in the primer binds to the guanine in the template. In the image, yellow only binds with red, and purple only binds with green, illustrating the principle of base pairing. <br> | ||
[[Image:121313.jpg]]<br>This picture illustrates the binding of the forward primer to the DNA template. Just as in the binding of the reverse primer to the DNA template, the binding of the forward primer to the DNA template occurs due to the attraction of complementary nucleotide bases.<br> | [[Image:121313.jpg]]<br>This picture illustrates the binding of the forward primer to the DNA template. Just as in the binding of the reverse primer to the DNA template, the binding of the forward primer to the DNA template occurs due to the attraction of complementary nucleotide bases.<br> | ||
[[Image:121314.jpg]]<br>After the forward and reverse primers have bound to separate strands of the DNA template, the DNA polymerase is ready to synthesize new DNA strands complementary to the template strands. <br> | [[Image:121314.jpg]]<br>After the forward and reverse primers have bound to separate strands of the DNA template, the DNA polymerase is ready to synthesize new DNA strands complementary to the template strands. <br> |
Revision as of 16:57, 29 October 2013
BME 100 Fall 2013 | Home People Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3 Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6 Course Logistics For Instructors Photos Wiki Editing Help | |||||||||||||
OUR TEAMLAB 1 WRITE-UPInitial Machine TestingThe Original Design
When we unplugged (part 3) from (part 6), the LCD screen on the machine remained lit, but did not display any information on the screen. When we unplugged the white wire that connects (part 6) to (part 2), the machine
We first tested the Open PCR machine on October 23, 2013. During this test run we had unsuccessful result, as our Open PCR machine failed to operate. ProtocolsThermal Cycler Program 1. Denature for one cycle at 95C for three minutes (Initial hold)
3. Extend the DNA at 72C for 3 minutes (to stabilize the DNA) DNA Sample Set-up
DNA Sample Set-up Procedure
Research and Development'PCR - The Underlying Technology' 'Components of a PCR Reaction'
'Steps of Thermal Cycling'
Thermal cycling initial starts at 95°C for three minutes during this period of time the enzymes are activated due to the optimal temperature that taq
polymerase will become active. During this step the single stranded DNA template will also begin to disconnect. The next step is Denature which will be at
95°C for 30 second throughout this time period the hydrogen bonds with the complementary bases will begin to become distorted ultimately causing the DNA
melting of the DNA template producing a single-stranded DNA molecule.
Following denaturation, the DNA undergoes annealing, where primers form stable DNA-DNA hydrogen bonds with the template DNA. After the template DNA
and the primers have bound, the DNA polymerase attaches to the primer-template hybrid. Annealing occurs at 57 degrees Celsius for 30 seconds.
Then, extension occurs at 72 degrees Celsius for 30 seconds. During extension, the DNA polymerase enzyme synthesizes a new strand of DNA complementary
to the template strand by adding dNTPs complementary to the bases of the template strand in the 5' to 3' direction.
During the final step which occurs at 72 degrees Celsius for duration of three minutes any remaining single stranded DNA that is leftover will become fully extended.
Also during the final step, the final hold occurs at four degree Celsius, which is when the entire reaction becomes stored.
'Diagram of PCR'
|