PCR - Electrophoresis Adding primers to the DNA for the PCR process.

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Presentation transcript:

PCR - Electrophoresis Adding primers to the DNA for the PCR process.

PCR - Electrophoresis A pipettor adds specific amounts of DNA polymerase, nucleotides, primers, and the templated DNA.

PCR - Electrophoresis The PCR machine is used to make copies of specific DNA sequences. This can be used to determine if a sample contains a specific gene or increase the number of copies of a gene for use in research.

PCR - Electrophoresis An electrophoresis machine. An agarose gel is place inside. Liquid covers the gel to prevent it from drying out and to allow the electricity to move through the gel.

PCR - Electrophoresis Gels are made from agarose. The agarose is melted in ddH2O and poured into the trays. Combs are placed in the trays to make the wells for the DNA to be injected into.

PCR - Electrophoresis The gel is allowed to cool. When they have solidified, they have the consistency of thick jello.

PCR - Electrophoresis Keeping track of the different gel recipes with post-it notes.

PCR - Electrophoresis The extracted DNA must be prepared for electrophoresis.

PCR - Electrophoresis A restriction enzyme is added to the DNA sample to cut it into smaller fragments.

PCR - Electrophoresis Blue dye is added to the DNA samples so they are easy to see while working with them. To add the dye, small droplets are placed on a non-absorbent surface.

PCR - Electrophoresis A DNA sample is pipetted from a microfuge tube onto one of the droplets of blue dye. The liquids are mixed by repeatedly pumping the them in & out with the pipettor.

PCR - Electrophoresis An electrophoresis machine with a agarose gel.

PCR - Electrophoresis The DNA samples are loaded into the wells of the electrophoresis machine with the pipettor.

PCR - Electrophoresis The DNA samples are loaded into the wells of the electrophoresis machine with the pipettor.

PCR - Electrophoresis The DNA samples are loaded into the wells of the electrophoresis machine with the pipettor. This takes a steady hand.

PCR - Electrophoresis The DNA samples are loaded into the wells of the electrophoresis machine with the pipettor.

PCR - Electrophoresis The machine is hooked up to a power source with a positive charge at one end and negative at the other.

PCR - Electrophoresis DNA has a slightly negative charge so it moves through the gel from negative to positive. (opposites attract)

PCR - Electrophoresis The gel must run for _____ hrs. or until you see the blue dye of the DNA samples near the end of the gel.

PCR - Electrophoresis The gel is taken from the electrophorsis machine and place on a UV light box. Ethidium bromide stains the DNA fragments so they glow under UV light.

PCR - Electrophoresis The gel on a light box. With normal light on, only the blue dye mixed in with the DNA can be seen.

PCR - Electrophoresis The gel on a light box. With normal light on, only the blue dye mixed in with the DNA can be seen.

PCR - Electrophoresis The gel on a light box. With normal light on, only the blue dye mixed in with the DNA can be seen.

PCR - Electrophoresis The gel on a light box. With normal light on, only the blue dye mixed in with the DNA can be seen.

PCR - Electrophoresis Once the normal lights are shut of, the fluorescing DNA stained with ethidium bromide can be seen. Each band is composed of DNA of the same length.

PCR - Electrophoresis Shorter DNA fragments travel through the gel faster because they cause less resistance in the gel. Therefore, they are found toward the farthest end of the gel.

PCR - Electrophoresis Large fragments are bigger & slower and only move a short distance through the gel.

PCR - Electrophoresis Electrophoresis gels are used to aid in a variety of things including gene mapping and crime investigation like the OJ Simpson trial.

PCR - Electrophoresis Computer analysis of the electrophoresis data. The data is sent to a professional company which determines the actual sequence of nucleotides in the DNA sample.

PCR - Electrophoresis Another representation of the nucleotide base pairs in the DNA sample.