BME100 s2018:Group9 W0800 L4: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Annaavila (talk | contribs)
Annaavila (talk | contribs)
Line 72: Line 72:
'''DNA Sample Set-up Procedure'''
'''DNA Sample Set-up Procedure'''
<!-- In your own words, write an easy-to-comprehend list of the steps your group took to set up the PCR reaction in the PCR reaction tubes. End with placing the tubes in to the thermal cycler. Do not copy-paste the instructions from the Workbook. That will be considered plagiarism. -->
<!-- In your own words, write an easy-to-comprehend list of the steps your group took to set up the PCR reaction in the PCR reaction tubes. End with placing the tubes in to the thermal cycler. Do not copy-paste the instructions from the Workbook. That will be considered plagiarism. -->
# Step 1: Extract DNA from cells.
* Step 1: Extract DNA from cells.
# step 2: Place the DNA into a special PCR tube.
* step 2: Place the DNA into a special PCR tube.
# Step 3: Add primer 1 to the PCR tube.
* Step 3: Add primer 1 to the PCR tube.
# Step 4: Add primer 2(will attach to the second site)
* Step 4: Add primer 2(will attach to the second site)
# Step 5: Add nucleotide to the PCR tube.
* Step 5: Add nucleotide to the PCR tube.
# Step 6: Add DNA Polymerase to the PCR tube.
* Step 6: Add DNA Polymerase to the PCR tube.
# Step 7: Place PCR tube, with all components, into a DNA thermal cycler.
* Step 7: Place PCR tube, with all components, into a DNA thermal cycler.


'''OpenPCR program'''
'''OpenPCR program'''

Revision as of 16:22, 14 March 2018

BME 100 Spring 2018 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 TEAM

Name: Anna
Avila
Name: Taylor
Dinubilo
Name: Tyler
Thompson
Name: Zoey
Wirtes
Name: student
Role(s)
Name: student
Role(s)

LAB 4 WRITE-UP

Protocol

Materials

  • Lab coat and disposable gloves
  • PCR reacon, mix, 8 tubes, 50 μL each: Mix contains Taq DNA polymerase, MgCl2, and dNTP’s

(hp://www.promega.com/resources/protocols/product‐informaon‐sheets/g/gotaq‐colorless ‐master‐mix‐m714‐protocol/)

  • DNA/ primer mix, 8 tubes, 50 μL each: Each mix contains a different template DNA. All tubes have the same forward primer and reverse primer
  • A strip of empty PCR tubes
  • Disposable pipee

ps: only use each only once. Never reuse disposable pipee ps. If you do, the samples will become cross‐contaminated

  • Cup for discarded ps
  • Micropipeor
  • OpenPCR machine: shared by two groups


PCR Reaction Sample List

Tube Label PCR Reaction Sample Patient ID
G9 + Positive control none
G9 - Negative control none
G9 1-1 Patient 1, replicate 1 44954
G9 1-2 Patient 1, replicate 2 44954
G9 1-3 Patient 1, replicate 3 44954
G9 2-1 Patient 2, replicate 1 75503
G9 2-2 Patient 2, replicate 2 75503
G9 2-3 Patient 2, replicate 3 75503


DNA Sample Set-up Procedure

  • Step 1: Extract DNA from cells.
  • step 2: Place the DNA into a special PCR tube.
  • Step 3: Add primer 1 to the PCR tube.
  • Step 4: Add primer 2(will attach to the second site)
  • Step 5: Add nucleotide to the PCR tube.
  • Step 6: Add DNA Polymerase to the PCR tube.
  • Step 7: Place PCR tube, with all components, into a DNA thermal cycler.

OpenPCR program

  • HEATED LID: 100°C
  • INITIAL STEP: 95°C for 2 minutes
  • NUMBER OF CYCLES: 25
  • Denature at 95°C for 30 seconds, Anneal at 57°C for 30 seconds, and
  • Extend at 72°C for 30 seconds
  • FINAL STEP: 72°C for 2 minute






Research and Development

PCR - The Underlying Technology






SNP Information & Primer Design

Background: About the Disease SNP


Primer Design and Testing