a spectrophotometer can be used to determine DNA quantity and quality In nucleic acids,

conjugated double bonds in purines and pyrimidines rings have a specific absorbance at 260nm,
which is proportional to their concentration (Beer-Lambert law) Therefore, the concentration of DNA
can be measured at absorbance A260 and is determined by performing the following calculation:

dsDNA concentration = A260 x conversion factor (47µ/ml) x dilution factor

Maximum protein absorption occurs at A 280 and ratio A260/A280 measures protein contamination in
the sample. In a buffered solution, the optimum value for pure dsDNA has an A 260/A280 of 1.85-1.88

It has been reported that absorbance at 260nm between 0.1 and 1.0 corresponds to reliable and
reproducible values Samples with low DNA concentration can be strongly affected by dust particles
or chemicals on laboratory microtubes that absorb UV light at 260nm and interfere with DNA
quantification The pipetting error can also reduce DNA concentration If a lower volume were
pipetted into the cuvette, it would appear as if a lower DNA volume was extracted.

More sensitive fluorescence methods are recommended if there is insufficient DNA concentration to
quantify by a spectrophotometer or DNA solution is contaminated

The minisatellite locus D1S80 is in the sub-telomeric, noncoding region of chromosome 1 which is
not associated with the disease phenotype The core sequence had 16 nucleotide base pairs, and the
total range of repeat lengths is approximately 370 to 802 bp The small size of D1S80 locus makes it
suitable for PCR analysis

Every participant will have received half of their markers from the mother and half from the father
Under the influence of applied voltage, negatively charged DNA molecules will travel through an
agarose matrix towards an anode with different velocities.
Practical 3

This practical aimed to examine restriction digests of genomic, plasmid and bacteriophage DNA by
gel electrophoresis.

Digestion products are separated by agarose gel according to their molecule length When placed in
the electric field, the negatively charged phosphorous backbone of DNA enables its migration
through a complex network of agarose pores towards the positively charged anode Vibrio fischeri is
a complex 4,284,050 bp genome that produced a bright band of unseparated DNA fragments at the
top of the gel due to the size limitation of agarose gel Pulse field gel electrophoresis is an alternative
and more complex method for separating fragments larger than 25kb pairs

Practical 4

DNA sequencing is a rapid procedure that could be applied in the design of effective diagnostic tools
such in FISH or PCR methods. Sequencing can also be employed to determine genetic diseases and
new disease causing agents

the ABI gene analyser, which separates sequenced materials according to their sizes and reads the
sequence. Then BLAST server was used to identify the nature of the DNA insert.