Automating a RAPD PCR Method for Analyzing Twin DNA Ppt Powerpoint Presentation slides-Medical Slideshow Statistical methods for analyzing DNA microarray data

Automating a RAPD PCR Method for Analyzing Twin DNA

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Slide 1 :

Automating a RAPD-PCR Method for Analyzing Twin DNA Dwayne Kalinay Kylie Rosengrant 28 April 2006

Slide 2 :

Background Information(Twins) 2 Categories of twin Monozygotic Dizygotic Fertilization Process

Slide 3 :

Polar Body twinning Fertilization Process Simultaneous fertilization Role of Polar Body 50-75% identical http://faculty.southwest.tn.edu/rburkett/A&P2_reproductive_system_lab.htm

Slide 4 :

Processes DNA Extraction Swabbing of human subjects Procedure Includes Solution Heat baths Vortexing

Slide 5 :

RAPD-PCR Master Mix (per Rxn) H20 PCR Buffer Mg dNTP P-13 Primer Taq DNApolymerase 11µl total of Master Mix Procedure Vortex Master Mix Add DNA template Vortex Add mineral oil Run in Thermocycler

Slide 6 :

Nature of P-13 Known as OPP-13 (from Operon Tech.) RAPD primer labeled with WELLRED D4, a fluorescent dye Dye fluoresces with the use of diode rays Specifically designed for CEQ genetic analysis systems

Slide 7 :

RAPD-PCR (continued) 2% Agarose Gel 1.2% FlashGel – checkpoint Procedure H20 each well mixed sample Includes Master Mix sample and dye Run @ 270 Volts QIAquick® PCR Purification Kit http://www.eastport.cz/firmy.php?kat1=bma&kat2=flashgel&kat3=index

Slide 8 :

DNA Sequencer Preparation for DNA Sequencer Size standard SLS (Sample Loading Solution) sample Capillary Electrophoresis Separation technique involving electric field Separates molecules based on: Size Charge Hydrophobicity

Slide 9 :

Results RAPD-PCR 2% Agarose gel smearing occurred FlashGel runs received strong banding Bands symbolize partial success in development of protocol

Slide 10 :

Pictures of FlashGel

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	SlidesOnline 1 Years ago. Category: other

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H20; PCR Buffer; Mg; dNTP; P-13 Primer; Taq DNApolymerase; 11µl total of Master Mix RAPD-PCR. 2% A more H20; PCR Buffer; Mg; dNTP; P-13 Primer; Taq DNApolymerase; 11µl total of Master Mix RAPD-PCR. 2% Agarose gel smearing occurred; FlashGel runs received less

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