1.Why does DNA move through an agarose gel?

Movement through the gel occurs when an electric current is applied across the gel. Since the gel is immersed in buffer, the current will travel through the bu ffer and gel, carrying the negatively charged DNA with it toward the positive anode 2.What are the two techniques used to create a DNA profile in this experiment? W hat function does each perform? a) Restriction fragment length polymorphism (RFLP) In RFLP, the DNA is cut into segments of varying lengths by an enzyme, then the segments separated out on the basis of size using a technique called electro phoresis. Fragments of a particular length are transfered to a nylon membrane. They ar e matched up with radioactively labelled fragments of DNA in such a way that only fragmen ts that are identical stick together. The excess radioactive fragments are washed away a nd an x-ray of the remaining fragments taken. This gives a picture of which of the label led fragments were in the original sample. b) Polymerase Chain Reaction (PCR) Amplify DNA sequences. c) Short tandem repeat profiling (STR) An enzyme is used to make many copies of a small section of the DNA. This se ction cut into pieces by another enzyme, and separated by electrophoresis. The fragmen ts are then visuallised with a silver stain, with the pattern of light and dark bands se en being characteristic for an individual 3.What is an Allele Ladder?What is its function in DNA profiling? Allele Ladder comprised of DNA fragments that represent common alleles at a l ocus.DNA Allele Ladder represents all the possible alleles at the BXP007 locus This is a reference, or marker, that you can compare your PCR reactions to so you can judge their relative sizes and their identities. These are the standard sizes of all the alleles known to occur at this locus. 4.What is required to visualize DNA following electrophoresis? Ethidium Bromide is used. This visualization technique, ethidium bromide is mixed with agarose powder, EDTA buffer and water to form the gel matrix before electrophoresis. As a result, the ethidium bromide molecules become uniformly dispersed throu ghout the matrix. Once the gel's wells have been filled with their respective DNA samples and t racking dyes, bases of the DNA molecules temporarily bind to the ethidium bromide particles . In the presence of ultraviolet light, ethidium bromide exhibits fluorescence. Then we shine a specially-calibrated UV light across the gel while a machine

captures the picture of the glowing fragments