PROCEDURE

By spinning laboratory samples

at very high speeds, the
components of a given mixture
are subjected to centrifugal
force, which causes more
dense particles to migrate away
from the axis of rotation and
lighter ones to move toward it.
These particles can sediment at
the bottom of the tube into
whats known as a pellet, and
this isolated specimen, or the
remaining solution, the
supernatant, can be further
processed or analyzed.
CENTRIFUGATION
is a process which involves the
application of the centripetal
force for the sedimentation of
heterogeneous mixtures with
a centrifuge, and is used in
industrial and laboratory
settings.
A centrifuge is a device that
spins liquid samples at high
speeds and thus creates a
strong centripetal force
causing the denser materials
to travel towards the bottom of
the centrifuge tube more
rapidly than they would under
the force of normal gravity.
 Centrifuge
A centrifuge is a machine designed to separate heavy material from
light material. When the substance is a very fine solid in a solution,
separating parts of the mixture can be
more difficult than a solid. In our case, we
will use a centrifuge to separate DNA from
a liquid so we can focus just on the DNA.
A centrifuge works by spinning the
substance at high speed. You have
probably seen the trick where you fill a
bucket of water about halfway full, spin it
around, and watch as the water stays in
the bucket. The water is pushed
against the bottom of the bucket by what is colloquially known as
centrifugal force. Though not a real force, but a reaction to the
centripetal force, centrifugal force pushes against the bottom of the
bucket. This causes the particles in the solution to clump at the bottom
of the tube. That solid clump is called a pellet, and the solution above it
is called the supernatant.
Here is a picture of a modern centrifuge. Notice the
different parts of the machine: there are holes for centrifuge tubes to be
inserted, the numbers of which vary machine to machine; the cap, to
secure the tubes inside, preventing them from flying out at high RPMs
(Revolutions Per Minute); the lid, which further protects both the samples
and the experimenter should something go wrong; a way to control the
RPM; and finally an on/off switch. Although RPM is commonly used for
setting centrifuge speeds, a more accurate measurement is actually
called RCF or relative centrifugal force. 1 RCF is independent of the radius
between the center of the centrifuge and the holes for the samples.
To use a centrifuge, put a test tube in one of the slots. However, it is
very important that the centrifuge be balanced! If the centrifuge is not
balanced the centrifuge will be unstable, and it will not spin at the right
speed! To balance a centrifuge, put samples in the centrifuge so they are
directly across from each other. If you only have one tube to centrifuge,
you should balance it with a blank tube (a tube filled with water). If
you have three tubes, you should put them in 1-7-13, so on and so forth.
Once the centrifuge is balanced, make sure the cap is screwed on tightly,
close the lid, set the desired RPM and time, and click start.