Microarrays

- The set of RNA molecules expressed in a cell at any

given time is called a transcriptone.
- Microarray technology allows the study of transcriptions
in real time. The technology exploits the basic principle
of hybridising complementary sequences.
- Microarrays are used to compare the genes present in
two species or to measure gene expression.
- Microarrays are based on a small piece of glass or
plastic. Short lengths of singles-stranded DNA are
attached to this support in a regular two-dimensional
pattern. Each individual position has multiple copies of
the same DNA probe.
- Steps of microarray use to compare the genomes of two
different species:
1. Select the DNA probes required and use an
automated process to apply those probes to the
positions on the miscarry.
2. DNA from the species is collected and denatured to
give single-stranded DNA.
3. The DNA is labeled with fluorescent tags, so that the DNA from the two different species
has different colours.
4. The labelled DNA samples are mixed together and allowed to hybridise with the DNA
probes on the microarray.
5. Any DNA that does not bind to the probes on the microarray is washed off.
6. The microarray is inspected with UV light, which causes the tags to fluoresce. Where the
microarray fluoresces hybridisation has occurred because the DNA fragments were
complementary to the probes.
Where there are green fluorescent spots there is DNA of one species, red the other, and
yellow both. Only the dark or blue spots have no hybridisation (not present in either
species).
Scan the microarray and store the data in a computer.
- Steps of microarray use to find out which genes are active in a cell at a particular time.
7. Gene probes from all of the genes in the human genome
are placed on the microarray.
8. The mRNA from the cytoplasm of a cell is isolated and
converted into single-stranded cDNA by reverse
transcriptase using DNA nucleotides that have fluorescent
tags.
9. We increase the number of DNA strands by PCR and then
the cDNA is applied to the microarray.
10. cDNA will hybridise with the appropriate fine probes, which
are detected when the microarray is scanned. The
fluorescent spots will indicate the genes that were
transcribed.
The higher the intensity of light from the spot, the more
that gene is being expressed.

Bioinformatics
- Bioinformatics combines biological data with computer
technology and statistics. It builds databases which can contain
gene sequences, the sequences of complete genomes, animo
acid sequences of proteins, and protein structures. All this can
be accessed via internet.
- Examples of databases
Ensembl: holds data on genomes of eukaryotic organisms.
UniProt (universal protein resource): holds information on the
primary sequences of proteins and the functions of many
proteins.
BLAST (basic local alignment search tool): An algorithm for
 comparing primary biological sequence information, such as the primary sequences of
different proteins or the nucleotide sequences of genes. researchers use BLAST to find
similarities between sequences that they are studying and those already saved in databases.
- Uses of the information from bioinformatics:
Closer similarity of biological sequences between species means more recent common
ancestry.
Human genes may be found in other organisms, such as Drosophila. Drosophila can be used
to investigate the way such genes have effects. Microarrays can be used to find out where
and when that gene is expressed during the development of the fruit fly.
Caenorhabditis elegans was the first multicellular organism to have its genome fully
sequenced. It was transparent, allowing the developmental fate of its cells to be mapped. It is
used as adel organism for studying the genetics of organ development due to its simplicity.
This provides information on the development of neurones, cell death, ageing, and behaviour.
Information about the genome of Plasmodium can now be used to find methods of controlling
the parasite. These methods may include vaccines for malaria.