Polymerase Chain Reaction (DNA Polymerase – duplicates DNA when cells divide) DNA copying machine – creates the compliment strand (ATCG-TAGC) PCR is used to amplify a short, well-defined part of a DNA strand. This can be a single gene, or just a part of a gene Requires certain components: DNA template, contains the region of the DNA fragment to be amplified Two primers, determines beginning and end of the region to be amplified DNA-Polymerase, which copies the region to be amplified Nucleotides, from which the DNA-Polymerase builds the new DNA Buffer, chemical environment for the DNA-Polymerase Thermocycler – amplification machine

detection of hereditary diseases the id of genetic fingerprints the cloning of genes paternity testing.

*Note: Each gel chamber holds 25 ml of the Agarose solution.* 1. Insert the comb and then tape ends of gels with scotch tape. Label the tape with your name. 2. Place gel tray into a clean weigh boat to contain spills. 3. Add 22.5 ml of dH 2 0 and 0.25g of Agarose to a 125 ml Erlenmeyer flask. 4. Microwave until it turns clear and all Agarose is in solution. (roughly 40 seconds-CAUTION: It bubbles quickly so do 10 second intervals) 5. Add 2.5 ml of 10x TAE buffer, then add 20  l ethidium bromide (EtBr). 6. Gently pour solution into gel tray, remove bubbles and let it sit for 20 minutes.

Agarose gel electrophoresis is a procedure that consists of injecting DNA into agarose gel and then applying an electric current to the gel. As a result, the smaller DNA strands move faster than the larger strands through the gel toward the positive current. The size of the PCR product can be determined by comparing it with a DNA ladder, which contains DNA fragments of known size, also within the gel

Our “dye” (EtBr) is added before

-Fill chamber with 2500 ml of 1x TAE -Put gel tray into chamber -Add dye to the extracted/digested DNA -dye weighs the DNA (dye + glycerol) -Load 1 st well with 15  l DNA Marker -Load remaining wells with 15  l dye/DNA mix Gene Ruler 1kb

Smaller fragments migrate faster DNA Is (+)

3 kb DNA Fragment