BME100 s2014:T Group6 L5
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
ProcedureSmart Phone Camera Settings
Solutions Used for Calibration
Placing Samples onto the Fluorimeter 2.) Second, place 80 microliter drop of SYBR Green 1 on the slide then place 80 microliter of the sample over the SYBR Green. 3.) Third, take a photo of the fluorimeter after lowering the lid of the box, so that it removes as much of stray light. 4.) Finally, remove the box without moving the smartphone and record the sample. Repeat for all samples.
Data AnalysisRepresentative Images of Samples Image J Values for All Samples
PCR Results Summary A polymerase chain reaction (PCR) was utilized to amplify the DNA from 2 patients identified here as 46296 and 16946. A positive and negative control were amplified alongside the patient’s DNA. PCR results were analyzed by using SYBR Green and a blue light emitting diode (LED) capable of causing the SYBR to fluoresce if DNA was present in the sample. Before the patient samples could be analyzed, known concentrations of calf thymus DNA had to be used in conjunction with the SYBR to create a curve fit correlating the calf thymus DNA concentration to the intensity of fluorescence. An iPhone 5 was used to take snap shots of 160 μL droplets, consisting of 80 μL of calf thymus solution that ranged from 0 – 5 μg/mL and 80 μL of SYBR Green. The photos were imported into Image J, a public, java based program available through the National Institute of Health (NIH). Image J has a function that calculates the integrated pixel density of image samples. The integrated density (RAWINTDEN) of the samples and the known DNA concentrations were incorporated in Graph 1 above.
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