AMPLIFYING DNA A.Recombinant DNA B.Polymerase Chain Reaction (PCR) (animation)

USING GEL ELECTROPHORESIS It is used to separate macromolecules like nucleic acids and proteins on the basis of size and charge. Gel electrophoresis is the movement of charged particles under the influence of an electric field.

A. USING NON-CODING DNA A lot more variability in non-coding DNA have alleles that have different numbers of random repeats EXAMPLE ONE INDIVIDUAL: TTAAGGTTAAGGTTAAGGTTAAGGTTAAGG TTAAGGTTAAGG METHODS FOR ESTABLISHING PATTERNS IN DNA

DNA fragments are made using restriction enzymes cutting the DNA strand at specific locations. Depending on where they cut, they form different length segments that migrate up to certain points on across the gel. Shorter fragments migrate the furthest distances. Coloured bands form showing where the segment has travelled to and the length of the segment can be determined. Each individual will produce a unique pattern or DNA fingerprint. B. USING RESTRICTION ENZYMES

EXAMPLE USE TTCC RESTRICTION ENZYME DNA SAMPLE ONE: ACTGTTCCACACTTCCGCACATCTTAATTCC

C. SANGER TECHNIQUE When replicating DNA segments, use regular nucleotides and dideoxynucleotides for the construction of the new DNA segment. Dideoxynucleotides (dd-A, dd-T, dd-C, dd-G) lack a 3’ OH group on the deoxyribose therefore if it is added to the growing DNA segment, then another nucleotide can not be attached to that segment and therefore terminating the segment prematurely. Fragments of the DNA segment are made varying in length. Gel electrophoresis is used to separate the fragments according to size. The bands indicate where a certain nucleotide is located on the segment of DNA. To analyze the entire sequence, you can run parallel gels using dd-A, dd-T, dd-C, and dd-G. A SOUTHERN BLOT is a process used to move DNA segments from a gel solution to a filter sheet. There is also Western, Northern, and Eastern Blots

TEMPLATE T C G C A T A G C C

STRAND A G C G T A T C G G A G C T

SOUTHERN BLOTTING