BME100 f2013:W1200 Group1 L4
|Line 78:||Line 78:|
| Tube A: Positive Control: cancer DNA template || Tube B: Patient 1 ID-16946 Replicate 1 || Tube C: Patient 1 ID-16946
| Tube A: Positive Control: cancer DNA template || Tube B: Patient 1 ID-16946 Replicate 1 || Tube C: Patient 1 ID-16946 Replicate 2 || Tube D: Patient 1 ID-16946 Replicate 3
| Tube E: Negative Control: non-cancer DNA template || Tube F: Patient 2 ID-46296 Replicate 1 || Tube G: Patient 2 ID-46296
| Tube E: Negative Control: non-cancer DNA template || Tube F: Patient 2 ID-46296 Replicate 1 || Tube G: Patient 2 ID-46296 Replicate 2 || Tube G: Patient 2 ID-46296 Replicate 3
Revision as of 15:42, 23 October 2013
|BME 100 Fall 2013|| Home |
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
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LAB 1 WRITE-UP
Initial Machine Testing
The Original Design
The PCR machine is heavily dependent on numerous connections and and interconnected parts. This means that, in order for it to work well, everything must be in its proper location and plugged in. This being said, when we unplugged part 3 (the LCD screen) from part 6 (circuit board) of the machine, the connection from the LCD to the circuit board was severed, resulting in the screen turning off. Also, when we unplugged the white wire that connects part 6 (circuit board) to part 2 (core) of the machine, the temperature changed from 26.7 degrees Celsius to -40 degrees Celsius. It is very important that all connections are correct in order to have an effective PCR machine.
Thermal Cycler Program
95°C for 3 minutes: The initial Template DNA strand separates
95°C for 30 seconds: The DNA double helix separates, creating two single-stranded DNA molecules
57°C for 30 seconds: The two primers attach to opposite ends of the top and bottom strand of the target DNA segment
72°C for 30 seconds: The DNA polymerase activates. Replication of the target DNA segment begins.
72°C for 3 minutes: The complementary binding of nucleotides continues until it gets o the end of the DNA strand and falls off.
4°C: The PCR reaction ends.
The PCR reaction mix will contain 8 tubes. Each tube will contain 50μL of the following:
The DNA primer mix contains 50μL of the following:
Research and Development
PCR - The Underlying Technology
(Add a write-up, essay-style, organized into paragrpahs with descriptive headers, based on the Q&A's from Section three of your worksheet)
(BONUS points: Use a program like Powerpoint, Word, Illustrator, Microsoft Paint, etc. to illustrate how primers bind to the cancer DNA template, and how Taq polymerases amplify the DNA. Screen-captures from the PCR video/ tutorial might be useful. Be sure to credit the sources if you borrow images.)