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https://drive.google.com/open?id=1MtEkNOjXXgC5-OFQ3OZxuxJ1UZbkVI-BwMGL7sKNh2g

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https://drive.google.com/open?id=1_Ul4IuMQEA_atsbt_mFUolkZBkV9jlSB

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What is the basic principle of gel electrophoresis?
Gel electrophoresis separates DNA fragments by size in a solid support medium (an agarose gel).
Why does DNA migrate toward positive electrode?
The DNA is negatively charged, so it run through the gel to the positive pole.

What is the TBE composition?
TBE is the buffer solution for the agarose gel, it's usually 10X and it has a 8.0 pH.

Why is important to add Mg ions? Are they co-enzymes or cofactors?
They are enzyme cofactors, and they are importante for the digestion

How can DNA be detected in agarose gel?
The DNA is transparent, so usually it's added some dye (in particular etherium bromide or cyber green).
Why are DNA dyes dangerous for human health?
The DNA dye can be dangerous because it's mutagen and a possible carcinogen, so use gloves.
What should be done to prepare an agarose gel? What should not be done preparing an agarose gel?
It's important mixing in intervals the solution for the agarose gel, and not let it in the microowave for too long without mixing. The flask must be covered with a loose cap, otherwise the pressure inside the container would become too high. The flask has to be also large enough to let the solution boil without coming it out. It's important using gloves for our safety, but also change them, in this way we will avoid contaminating other instruments or parts of the lab. The agarose has to be completely dissolve, or we will have regions with different concentrations. While the gel is setting, don't move the gel tray, or it can be damaged. Remove the comb slowly and with vertical motion

Which piece of information can be find out from a DNA pattern bands?
The fragments are separated by electrophoresis, and the different patterns of DNA fragments are compared (in our case the culprit and the suspects). The pattern with the same fragments dimension and position comes from the same person.

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What role does DNA evidence play in solving crimes? (YOUTUBE VIDEO)

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What role does DNA evidence play in solving crimes? (YOUTUBE VIDEO)

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Why has DNA received a  lot of media attention?
Because it has a fantastic power of discrimination. If we run a genotype and we compare it back to someone, the match probability is in the billions or trillions to one. It's an evidence stronger than eyewitness testimony.
How long has DNA been used to identify people?
The DNA is used to identify people since mid 80s.
The techniques used nowadays are more sensitive. How much DNA is needed to do a test?
Now you need less than a gram of DNA

How many cells are needed to get a full profile?
With three or four cells you can still get a full profile.

In what conditions must the exhibit be kept?
The DNA can be also fresh, but if it isn't in good conditions, for example in a wet room where the mold can grow, the DNA will degrade quickly.
How long does DNA last if taken in good conditions?
But if we freeze the DNA, this can last forever.

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the voltage applied to the electrophoretic cell, the buffer solution pH, the agarose gel concentration and the presence of bubble in the gel are the things that influence more gel electrophoreis results.
The first one because if the voltage is too high, the DNA can be destroyed.
The second because the pH makes the DNA negatively charged, in this way during the electrophoresis it will run to the positive pole.
The third because change the size of the molecular sieve.
The last one, so the bubble, would make the gel unclear, and the DNA wouldn't be able to run in a regular way.

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Double stand: doppio filamento

Dandruff: forfora

Teeth pattern: modello dei denti

Knighted: titolo di cavaliere

Landmark: punto di riferimento

Marker: marcatore

Head-tail: testa coda

Probe: sonda (o sondare come verbo)

Sheet lay: foglio di carta

Tray: vassoio

Loaded: caricato

Wells: pozzi

Peak: picco
Sieve: setaccio
Womb: grembo
Lumps: grumi
Sample: campione

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1 Was it possible to identify people in the past? How?
Yes, it was. In the past human identification has been based on finger-printing, since 1901; but the result of this investigation could demonstrate only if somebody had the same type of proteins of the culprit but there were million of people with the same pattern, so it could be only used to exclude from a crime and not to accuse anybody.
In 1901 blood typing groups were discovered by Karl Landsteiner but there are only four different types and lots of people can share the same type. This kind of marker could be used only to exclude: if the culprit was A blood type group and the suspected was of a different one, we could consider him innocent.
2 When was used first DNA finger printing?
This technique was developed in 1984 by Alec J. Jeffreys, but this method was used for the first time when he was asked to help in a disputed immigration case to confirm the identity of a British boy whose family was originally from Ghana. The case was resolved when the DNA results proved that the boy was closely related to the other members of the family.
3 Where can DNA be obtained from?
The DNA can be obtained from sperm, saliva (so mucosal surfaces), urine, blood (white blood cells), hair with roots, skin cells (es dandruff)
4 Who invented RFLP technique? When?
RFLP means restriction fragments length polymorphism. It based on different lengths of fragments produced by restriction enzymes digestion, analyzed by electrophoresis.
It's a technique invented in 1984 by Alec J. Jeffreys
5 What is a restriction enzyme?
Restriction enzymes are DNA-cutting enzymes found in bacteria.
6 What is restriction enzyme role in nature?
They are inside bacteria cytoplasm to prevent viral o bacteriophage infection. They cut in pieces double strand viral DNA injected by viral particles.