BME100 s2014:W Group5 L5: Difference between revisions
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'''Placing Samples onto the Fluorimeter''' | '''Placing Samples onto the Fluorimeter''' | ||
# '' | # ''On the first two rows of the slide, place one drop (the pipette is calibrated to 80 microlitres) of SYBR Green.'' | ||
# '' | # ''Add a second drop of the diluted calf thymus DNA on top of the first drop on the slide. We now have a sample!'' | ||
# '' | # ''Adjust the slide such that the blue LED light is using the drop to focus on the other side of the drop.'' | ||
# '' | # ''Use the camera's timer to take a picture after closing the light box (this prevents as much outside light as possible from contaminating our data).'' | ||
# ''Remove the box, being careful not to knock the smartphone out of position.'' | |||
# ''Use the pipette to remove the entire 160 microlitre drop from the slide, then move the slide into the next position and use the next concentration standard.'' | |||
# ''Repeat as needed.'' | |||
<br> | <br> | ||
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'''Image J Values for All Samples''' | '''Image J Values for All Samples''' | ||
'' | {| | ||
| align="center" style="background:#f0f0f0;"|'''PCR Product TUBE LABEL''' | |||
| align="center" style="background:#f0f0f0;"|'''Diluted Volume (µL)''' | |||
| align="center" style="background:#f0f0f0;"|'''SYBR GREEN Volume (µL)''' | |||
| align="center" style="background:#f0f0f0;"|'''INTDEN 1''' | |||
| align="center" style="background:#f0f0f0;"|'''INTDEN2''' | |||
'''Fitting a Straight Line'''<br> | | align="center" style="background:#f0f0f0;"|'''INTDEN3''' | ||
| align="center" style="background:#f0f0f0;"|'''AVERAGE INTDENS VALUE''' | |||
|- | |||
| A1||80||80||2716687||3229970||3340085||3095581 | |||
|- | |||
| A2||80||80||1451878||2568844||8446955||4155892 | |||
|- | |||
| A3||80||80||3862714||8011744||1600043||4491500 | |||
|- | |||
| B1||80||80||9856858||5058003||3108259||6007707 | |||
|- | |||
| B2||80||80||6862644||5685363||6783718||6443908 | |||
|- | |||
| B3||80||80||6287959||7119245||9598254||7668486 | |||
|- | |||
| +||80||80||11779932||7973600||11270538||10341357 | |||
|- | |||
| -||80||80||4371681||2482949||1977785||2944138 | |||
|- | |||
| | |||
|}'''Fitting a Straight Line'''<br> | |||
''Linear Regression of PCR Results'' | |||
<br> | |||
[[Image:Dna_calibration_graph.jpg]] | |||
<br> | <br> | ||
< | ''Interpretation''<br><br> | ||
Based on the results from the image analysis, the intensity in the green coloration in patient B was higher than patient A. Due to the fact that false positives and negatives were controlled for, it can be confidently said that patient B's DNA indicated a positive result for gene detection test. Patient A did not test positive. According to the calibration, it should be noted that the concentration of DNA is positively correlated to the green intensity of the SYBR Green dye; thus the high intensity in patient B's amplified DNA would be expected to increase with increased concentration of DNA. |
Revision as of 23:42, 15 April 2014
BME 100 Spring 2014 | 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 5 WRITE-UPBackground InformationSYBR Green Dye
ProcedureSmart Phone Camera Settings
First turn on the Blue LED switch in order to activate the excitation light. Also make sure the camera is open with the settings listed above. From there place the pone on the cradle with 90 degree angle from the slide. Make sure to adjust the height of phone and/or fluorimeter in order to take pictures of the drops. Lastly, the cradle should be as close to the fluorimeter as possible with having a clear picture still.
4.5 cm
Placing Samples onto the Fluorimeter
Data AnalysisRepresentative Images of Samples [Instructions: Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with no DNA] [Instructions: Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with DNA (positive signal)]
Image J Values for All Samples
Linear Regression of PCR Results
Interpretation |