BME100 s2014:T Group9 L5: Difference between revisions
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|- valign="top" | |- valign="top" | ||
| [[Image: | | [[Image:cassisicoolerthanyou.jpg|100px|thumb|Name: Cassaundra Kinnear<br>]] | ||
| [[Image: | | [[Image:meandbella.JPG|100px|thumb|Name: Katherine Underwood<br>]] | ||
| [[Image: | | [[Image:kylequeener.JPG|100px|thumb|Name: Kyle Queen<br>]] | ||
| [[Image: | | [[Image:Mohammed's Pic.JPG|100px|thumb|Name: Mohammed Binsheliail<br>]] | ||
|} | |} | ||
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'''Single-Drop Fluorimeter'''<br> | '''Single-Drop Fluorimeter'''<br> | ||
A single-drop fluorimeter is made up of a sturdy black cover used to block out the light. There is a phone cradle used o hold the camera while taking pictures. Inside the black cover there is a platform to hold the slides. There is also an LED light placed around the slides. This light emits a blue light used to see the SYBR green dye. | A single-drop fluorimeter is made up of a sturdy black cover used to block out the light. There is a phone cradle used o hold the camera while taking pictures. Inside the black cover there is a platform to hold the slides. There is also an LED light placed around the slides. This light emits a blue light used to see the SYBR green dye. | ||
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'''Smart Phone Camera Settings'''<br> | '''Smart Phone Camera Settings'''<br> | ||
* Type of Smartphone: iPhone 4s | |||
** Flash: off | |||
** ISO setting: N/A | |||
** White Balance: N/A | |||
** Exposure: N/A | |||
** Saturation: N/A | |||
** Contrast: N/A | |||
* Type of Smartphone: | |||
** Flash: | |||
** ISO setting: | |||
** White Balance: | |||
** Exposure: | |||
** Saturation: | |||
** Contrast: | |||
'''Calibration'''<br> | '''Calibration'''<br> | ||
<br> | |||
The camera was set on a cradle eye level with the sample. We set the camera with a timer so there would be no other light than the LED light the green glow coming from the sample. We clicked the button and closed the flap of the outer black case, then the camera took the picture of the sample. | |||
* Distance between the smart phone cradle and drop = 8 cm | |||
'''Solutions Used for Calibration' | '''Solutions Used for Calibration''' | ||
{| {{table}} width=700 | {| {{table}} width=700 | ||
|- | |- | ||
| | | Initial Concentration of 2X Calf Thymus DNA solution (micrograms/mL) || Volume of the 2X DNA solution (µL) || Volume of the SYBR GREEN I | ||
Dye solution (µL) | |||
|| Final DNA concentration in SYBR Green I solution (µg/mL) | |||
|- | |||
| 5 || 80 || 80 || 2.5 | |||
|- | |||
| 2 || 80 || 80 || 1 | |||
|- | |||
| 1 || 80 || 80 || 0.5 | |||
|- | |- | ||
| | | 0.5 || 80 || 80 || 0.25 | ||
|- | |- | ||
| | | 0.25 || 80 || 80 || 0.125 | ||
|- | |||
| 0 || 80 || 80 || 0 | |||
|} | |} | ||
'''Placing Samples onto the Fluorimeter''' | '''Placing Samples onto the Fluorimeter''' | ||
# '' | # ''Place the fluorimeter into the box'' | ||
# '' | # ''Place the camera in the cradle '' | ||
# '' | # ''Adjust the height of the fluorimeter with the trays so the camera will see the drop at eye level'' | ||
# '' | # ''Insert the slide into the fluorimeter'' | ||
# ''Adjust the distance between the camera and the slide to be as close as possible without being blury'' | |||
# ''Place an 80 microliters drop of SYBR Green in between the first two rows of the slide'' | |||
# ''Add 80 microliters of the indicated DNA solution to the drop of SYBR Green'' | |||
# ''Align the sample and the blue LED light'' | |||
# ''Click the timer on the camera and close the lid for the picture'' | |||
# ''Take one more picture, for a total of two pictures, without moving the camera'' | |||
<br> | <br> | ||
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==Data Analysis== | ==Data Analysis== | ||
'''Representative Images of Samples''' | '''Representative Images of Samples''' | ||
[[Image:Droplet_(2).png]] | |||
[[Image:Droplet_2_(2).png]] | |||
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'''Image J Values for All Samples''' | '''Image J Values for All Samples''' | ||
'' | {| | ||
| align="center" style="background:#f0f0f0;"|'''Concentration''' | |||
| align="center" style="background:#f0f0f0;"|'''Area''' | |||
| align="center" style="background:#f0f0f0;"|'''Mean''' | |||
| align="center" style="background:#f0f0f0;"|'''IntDen''' | |||
| align="center" style="background:#f0f0f0;"|'''RawIntDen''' | |||
| align="center" style="background:#f0f0f0;"|'''RawIntDen Back''' | |||
|- | |||
| 0||89824||22.134||1988142||1988142||4075684 | |||
|- | |||
| ||106024||25.339||2686504||2686504||3520178 | |||
|- | |||
| ||244420||39.829||9735070||9735070||4252614 | |||
|- | |||
| 0.25||232620||54.65||12712759||12712759||1470699 | |||
|- | |||
| ||213096||60.944||12986996||12986996||1545487 | |||
|- | |||
| ||230408||43.885||10111560||10111560||1460134 | |||
|- | |||
| 0.5||316088||86.678||27397838||27397838||5561052 | |||
|- | |||
| ||263648||111.592||29421095||29421095||5410757 | |||
|- | |||
| ||228141||134.47||30678127||30678127||1187005 | |||
|- | |||
| 1||275068||121.335||33375368||33375368||1276411 | |||
|- | |||
| ||285964||116.944||33441633||33441633||1345189 | |||
|- | |||
| ||99532||113.984||11345102||11345102||1352381 | |||
|- | |||
| 2||101468||114.739||11642363||11642363||921896 | |||
|- | |||
| ||101884||125.741||12810987||12810987||957122 | |||
|- | |||
| ||109756||115.55||12682322||12682322||1114350 | |||
|- | |||
| 5||108804||173.976||18929237||18929237||654847 | |||
|- | |||
| ||109360||120.332||13159480||13159480||796070 | |||
|- | |||
| ||106772||111.949||11952999||11952999||918322 | |||
|- | |||
| | |||
|} | |||
[http://excel2wiki.net/wikipedia.php Excel-to-Wiki Converter]. | |||
{| | |||
| align="center" style="background:#f0f0f0;"|'''Tube A''' | |||
| align="center" style="background:#f0f0f0;"|'''Tube B''' | |||
| align="center" style="background:#f0f0f0;"|'''Tube C''' | |||
| align="center" style="background:#f0f0f0;"|'''Tube D''' | |||
| align="center" style="background:#f0f0f0;"|'''Tube E''' | |||
| align="center" style="background:#f0f0f0;"|'''Tube F''' | |||
| align="center" style="background:#f0f0f0;"|'''Tube G''' | |||
| align="center" style="background:#f0f0f0;"|'''Tube H''' | |||
|- | |||
| 2308221||4130155||5069909||5325157||11018266||14630361||17579974||21359877 | |||
|- | |||
| 4078569||4773563||5282577||5753055||11153155||16780232||18268541||21671092 | |||
|- | |||
| 4107249||5055747||5291947||7644472||13454922||17148850||20260575||22388687 | |||
|- | |||
| | |||
|} | |||
'''Fitting a Straight Line'''<br> | '''Fitting a Straight Line'''<br> | ||
[[Image:Graph_(2).png]] | |||
<!-- Note: Be sure to delete the text in brackets: ''[ ]'' --> | <!-- Note: Be sure to delete the text in brackets: ''[ ]'' --> | ||
Latest revision as of 09:41, 17 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 TEAM
LAB 5 WRITE-UPBackground InformationSYBR Green Dye SYBR green dye is used as a nucleic acid stain and binds to DNA. The result is a DNA-dye-complex that absorbs blue light and emits green light. This dye can also bind to single-stranded DNA and RNA with a lower performance.
A single-drop fluorimeter is made up of a sturdy black cover used to block out the light. There is a phone cradle used o hold the camera while taking pictures. Inside the black cover there is a platform to hold the slides. There is also an LED light placed around the slides. This light emits a blue light used to see the SYBR green dye.
How the Fluorescence Technique Works The blue LED light shines on the DNA-dye samples. The amount of DNA in each sample is measured based on the concentration of the green light being emitted from the sample.
ProcedureSmart Phone Camera Settings
Solutions Used for Calibration
Placing Samples onto the Fluorimeter
Data AnalysisRepresentative Images of Samples
Image J Values for All Samples
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