BME100 s2014:W Group9 L5: Difference between revisions
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'''SYBR Green 1 Dye'''<br> | '''SYBR Green 1 Dye'''<br> | ||
SYBR Green 1 Dye is | SYBR Green 1 Dye is used as a nucleic acid stain and has numerous uses in the biology world. It is a small molecular dye that illuminates effectively while in the presence of double-stranded DNA, but does not illuminate as well when mixed with water or single-stranded DNA. The green dye can be placed in a sample with possible DNA and will glow a bright green if DNA is present. This is the most common use of the dye and helps qualitatively analyze if DNA is present in a substance. The dye can also be placed inside a PCR mixture and this allows the dye to bind to the single-stranded DNA. It can still be used with both, and additionally RNA. | ||
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'''Single-Drop Fluorimeter'''<br> | '''Single-Drop Fluorimeter'''<br> | ||
The Single-Drop Fluorimeter consists of a small, simple plastic box with two notches where the multi-welled slide can fit in. It has a switch located on the right hand side that allows for the blue LED light to be turned on or off. The slide of the fluorometer is aligned so that the blue LED shines through a drop placed over two middle wells. The slide has holes along it to allow for a sample to be placed in them. Placing the drops must be carefully done to ensure no overflow. | |||
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[[http://openwetware.org/images/7/71/Group4_Fluorometer.JPG]] | |||
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'''How the Fluorescence Technique Works'''<br> | '''How the Fluorescence Technique Works'''<br> | ||
The fluorometer has a slide with small holes where drops of sample DNA were placed. The slide layer is made of Teflon, giving the slide texture to allow the sample drop to maintain its "drop" shape. The reason this is necessary is because the light from the fluorometer can go directly through the drop to increase the intensity of the DNA glow. It also allows for the SYBR Green to move to the surface, which allows for the glow to be seen on the top of the drop. All of this makes it easier to observe and analyze whether the sample has a green glow, and whether there was DNA present in the sample. | |||
Revision as of 21:22, 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 TEAM
LAB 5 WRITE-UPBackground InformationSYBR Green 1 Dye Single-Drop Fluorimeter How the Fluorescence Technique Works
ProcedureSmart Phone Camera Settings
Calibration [Instructions: Describe how to set up your camera in front of the fluorimeter. Add a PHOTO of this set-up for bonus points.]
[Instructions: See worksheet page 6.]
[Add more rows as needed]
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 [Instructions: See worksheet page 8. To save time on typing a new Wiki table from scratch, use THIS TOOL to auto-generate a Wiki table: Excel-to-Wiki Converter. Copy the headers and values from the Excel spreadsheet you made, paste them into the form field, click submit, copy the Wiki code that the tool generated, and replace TABLE GOES HERE (below) with your auto-generated code.]
[Instructions: Place an IMAGE of your Excel plot with a line of best fit here. See worksheet page 9]
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