Creating a DNA Profile

If they're all supposed to arrive at a similar result -- a unique DNA profile -- then why are there
so many different techniques for analysis? Which technique to use depends on a couple of
factors, including cost, time available for analysis and the quality and amount of the DNA
sample available.

The first method for creating a DNA profile was RFLP, or restriction fragment length
polymorphism. RFLP is not used as often today because it requires a large sample of DNA -- as
much as 25 hairs or a nickel-sized spot of bodily fluid -- and can take as long as a month to
complete [source: Baden]. It also requires examining multiple sections of the DNA strand to find
variations, which is time-consuming and leaves more room for human error. Some of the steps
for RFLP analysis are also used in other types of DNA profiling. For RFLP, the steps are:
1. Separate white and red blood cells with a centrifuge.
2. Extract DNA nuclei from the white blood cells. This is done by bathing the cells in hot
water, then adding salt, and putting the mixture back into the centrifuge.
3. Cut DNA strand into fragments using a restriction enzyme.

4. Place fragments into one end of a bed of agarose gel with electrodes in it. Agarose gel is
made from agar-agar, a type of seaweed that turns into gelatin when dissolved in boiling
water.
5. Use an electric current to sort the DNA segments by length. This process is called
agarose gel electrophoresis. Electrophoresis refers to the process of moving the
negatively-charged molecules through the gel with electricity. Shorter segments move
farther away from their original location, while longer ones stay closer. The segments
align in parallel rows.
6. Use a sheet of nitrocellulose or nylon to blot the DNA. The sheet is stained so the
different lengths of DNA bands are visible to the naked eye. By treating the sheet with
radiation, an autoradiograph is created. This is an image on x-ray film left by the decay
pattern of the radiation. The autoradiograph, with its distinctive dark-colored parallel
bands, is the DNA profile.
PCR (polymerase chain reaction) analysis is usually the first step in the creation of a DNA
profile today. PCR can replicate a small amount of DNA to create a larger sample for analysis. It
does this using a repeating process that takes about five minutes. First, a heat-stable DNA
polymerase -- a special enzyme that binds to the DNA and allows it to replicate -- is added.
Next, the DNA sample is heated it to 200 degrees F (93 degrees C) to separate the threads. Then
the sample is cooled and reheated. Reheating doubles the number of copies. After this process is
repeated about 30 times, there is enough DNA for further analysis.
PCR is the first step in analyzing STRs (Short Tandem Repeats), which are very small, specific
alleles in a variable number tandem repeat (VNTR). Alleles are pairs of genes that occur
alternately at a specific point, or loci, on a chromosome. STRs are explained further in How
DNA Evidence Works. Analyzing STRs is more accurate than the RFLP technique because their
small size makes them easier to separate and to tell apart.
A variation on STR analysis is Y-STR. Only STRs found on the Y-chromosome (which only
males have) is analyzed. STR analysis is useful if the sample has mixed DNA (from both men
and women) or in sexual assault cases with a male assailant. Y-STR is otherwise processed just
like a regular STR.
AmpFLP, amplified fragment length polymorphism, is another technique that uses PCR to
replicate DNA. Like RFLP, it first uses a restriction enzyme. Then, the fragments are amplified
using PCR and sorted using gel electrophoresis. AmpFLP's advantage over other techniques is
that it can be automated and doesn't cost very much. However, the DNA sample must be high
quality or errors may result, which is the case with most DNA analysis techniques. Analysts can
have a time telling the longer strands apart because they bunch up tightly.
Eg: Human remains, goatskins, leather, pollen, fish scales, leaves, seeds, feathers, hair, blood,
textiles, wood, or (wool, silk, cotton, linen/flax)

Extract DNA

Use restriction enzyme(to break DNA into fragments)

Use polymerase chain reaction(to increase concentration of fragments)
Place sample on agrose gel
Carry out electrophoresis
Label fragments(transferred to a membrane) with radioactive isotope