BME103 s2013:T900 Group7 L2: Difference between revisions
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'''SYBR Green Dye | '''SYBR Green Dye '''<br> | ||
SYBR Green Dye is a membrane permeable nucleic acid dye used in molecular biology labs. 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. Although the DNA used in our samples was double stranded DNA, the SYBR Green Dye can also be used to detect single stranded DNA because it has such a high affinity. Although it was placed in the samples after being in the PCR machine, it also can be placed inside a PCR mixture and this allows the dye to bind to the single stranded DNA before it is synthesized by the Taq Polymerase. However, the result is the same. Either way it binds to DNA and glows when DNA is present. <br> | SYBR Green Dye is a membrane permeable nucleic acid dye used in molecular biology labs. 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. Although the DNA used in our samples was double stranded DNA, the SYBR Green Dye can also be used to detect single stranded DNA because it has such a high affinity. Although it was placed in the samples after being in the PCR machine, it also can be placed inside a PCR mixture and this allows the dye to bind to the single stranded DNA before it is synthesized by the Taq Polymerase. However, the result is the same. Either way it binds to DNA and glows when DNA is present. <br> | ||
[[Image:Shire_device.JPG|x400px|left|thumb|Single-Drop Fluorimeter]]<br> | [[Image:Shire_device.JPG|x400px|left|thumb|Single-Drop Fluorimeter]]<br> | ||
'''Single-Drop Fluorimeter'''<br> | '''Single-Drop Fluorimeter'''<br> | ||
The Single-Drop Fluorimeter consists of a small, simple plastic box with a apparatus situated on the top where the multi-welled slide can fit in | The Single-Drop Fluorimeter consists of a small, simple plastic box with a apparatus situated on the top where the multi-welled slide can fit in. It has a switch located on the right hand side that allows for the LED light to be turned on or off. The slide of the fluorometer is align 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. | ||
'''How the Fluorescence Technique Works | '''How the Fluorescence Technique Works'''<br> | ||
The fluorometer has a slide with small holes where drops of sample DNA were placed. The drops maintain their round shape because there is a Teflon layer on the slide. The Teflon gives the slide texture and allows 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. | The fluorometer has a slide with small holes where drops of sample DNA were placed. The drops maintain their round shape because there is a Teflon layer on the slide. The Teflon gives the slide texture and allows 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. | ||
Latest revision as of 10:20, 2 April 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 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
OUR TEAM
LAB 2 WRITE-UPBackground InformationSYBR Green Dye Single-Drop Fluorimeter How the Fluorescence Technique Works
ProcedureSmart Phone Camera Settings
Calibration
Placing Samples onto the Fluorimeter
Data AnalysisRepresentative Images of Samples
Image J Values for All Samples
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