BME103 s2013:T900 Group2

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Research and Development)
(Research and Development)
Line 82: Line 82:
Polymerase Chain Reaction (PCR) is a scientific method that utilizes DNA Polymerase to create a complimentary base strand from a template strand of DNA. Triphosphate nucleotides align with open DNA strands and DNA polymerase works to link the complementary nucleotide bases together growing strands through both condensation and hydroysis reactions. One major issue with DNA polymerase is that DNA strands are anti parallel, and polymerase is only able to add nucleotides to the free 3'OH group hence it can only build new strands in the 5'-3' direction. Therefore, to correct for this issue the presence of a primer is required so that polymerase can proceed with directing the new nucleotides in place. Through these mechanisms it is possible to target specific positions on the template DNA sequence that a scientist intends to amplify. When the PCR process is completed the targeted DNA sequence containing the single-nucleotide polymorphism (SNP) will have manufactured over a billion copies (amplicons).
Polymerase Chain Reaction (PCR) is a scientific method that utilizes DNA Polymerase to create a complimentary base strand from a template strand of DNA. Triphosphate nucleotides align with open DNA strands and DNA polymerase works to link the complementary nucleotide bases together growing strands through both condensation and hydroysis reactions. One major issue with DNA polymerase is that DNA strands are anti parallel, and polymerase is only able to add nucleotides to the free 3'OH group hence it can only build new strands in the 5'-3' direction. Therefore, to correct for this issue the presence of a primer is required so that polymerase can proceed with directing the new nucleotides in place. Through these mechanisms it is possible to target specific positions on the template DNA sequence that a scientist intends to amplify. When the PCR process is completed the targeted DNA sequence containing the single-nucleotide polymorphism (SNP) will have manufactured over a billion copies (amplicons).
-
The SNP included in the amplicon for this experiment was  denoted as rs17879961. This polymorphism is a variant of the CHEK2 gene which if present in a person's genome may increase the risk of developing breast cancer. This SNP is signified by a single base change from a Thymine (T) to a Cytosine(C) located on chromosome 22.
+
The SNP included in the amplicon for this experiment was  denoted as rs17879961. This polymorphism is a variant of the CHEK2 gene which if present in a person's genome may increase the risk of developing breast cancer. This SNP is signified by a single base change from a Thymine (T) to a Cytosine (C) located on chromosome 22.
(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 OpenPCR tutorial might be useful. Be sure to credit the source if you borrow images.)
(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 OpenPCR tutorial might be useful. Be sure to credit the source if you borrow images.)

Revision as of 17:38, 25 March 2013

BME 103 Spring 2013 Home
People
Lab Write-Up 1
Lab Write-Up 2
Lab Write-Up 3
Course Logistics For Instructors
Photos
Wiki Editing Help
Image:BME494_Asu_logo.png

Contents

OUR TEAM

Andy SonProtocol Planner
Andy Son
Protocol Planner
William Scott R&D Scientist
William Scott
R&D Scientist
Joe Sasnone R&D Scientist
Joe Sasnone
R&D Scientist
Name: studentRole(s)
Name: student
Role(s)
Name: studentRole(s)
Name: student
Role(s)
Name: studentRole(s)
Name: student
Role(s)

LAB 1 WRITE-UP

Initial Machine Testing

The Original Design
(Add image of the full OpenPCR machine here, from the Week 9 exercise. Write a paragraph description for visitors who have no idea what this is)


Experimenting With the Connections

When we unplugged (part 3) from (part 6), the machine ... (did what? fill in your answer)

When we unplugged the white wire that connects (part 6) to (part 2), the machine ... (did what? fill in your answer)


Test Run

(Write the date you first tested Open PCR and your experience(s) with the machine)




Protocols

Thermal Cycler Program

DNA Sample Set-up

row 1 cell 1 row 1 cell 2 row 1 cell 3 row 1 cell 4
row 2 cell 1 row 2 cell 2 row 2 cell 3 row 2 cell 4

DNA Sample Set-up Procedure

  1. Step 1
  2. Step 2
  3. Step 3...

PCR Reaction Mix

  • What is in the PCR reaction mix?

DNA/ primer mix

  • What is in the DNA/ primer mix?






Research and Development

Specific Cancer Marker Detection - The Underlying Technology

Polymerase Chain Reaction (PCR) is a scientific method that utilizes DNA Polymerase to create a complimentary base strand from a template strand of DNA. Triphosphate nucleotides align with open DNA strands and DNA polymerase works to link the complementary nucleotide bases together growing strands through both condensation and hydroysis reactions. One major issue with DNA polymerase is that DNA strands are anti parallel, and polymerase is only able to add nucleotides to the free 3'OH group hence it can only build new strands in the 5'-3' direction. Therefore, to correct for this issue the presence of a primer is required so that polymerase can proceed with directing the new nucleotides in place. Through these mechanisms it is possible to target specific positions on the template DNA sequence that a scientist intends to amplify. When the PCR process is completed the targeted DNA sequence containing the single-nucleotide polymorphism (SNP) will have manufactured over a billion copies (amplicons).

The SNP included in the amplicon for this experiment was denoted as rs17879961. This polymorphism is a variant of the CHEK2 gene which if present in a person's genome may increase the risk of developing breast cancer. This SNP is signified by a single base change from a Thymine (T) to a Cytosine (C) located on chromosome 22.

(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 OpenPCR tutorial might be useful. Be sure to credit the source if you borrow images.)

Sources:

Brennan, Paul et al. Uncommon CHEK2 Mis-sense Variant and Reduced Risk of Tobacco-related Cancers: Case-control Study. Rep. 15th ed. Vol. 16. Oxford: Advance Access, 2007. Human Molecular Genetics. Oxford University Press, 21 May 2007. Web. 25 Mar. 2013. PDF. <http://hmg.oxfordjournals.org/content/16/15/1794.full.pdf>.

"PCR Introduction." NCBI. U.S. National Library of Medicine, n.d. Web. 25 Mar. 2013. <http://www.ncbi.nlm.nih.gov/projects/genome/probe/doc/TechPCR.shtml>.

Sadava, David E. Life: The Science of Biology. Sunderland, MA: Sinauer Associates, 2011. Print.






Personal tools