Professional Documents
Culture Documents
CPTCB One Month Traning
CPTCB One Month Traning
CPTCB One Month Traning
BIOTECHNOLOGY
To prepare the MS- B2 or the BAP (benzenyl Amino Purine) medium for the plant tissue culture.
Principle:
For tissue culture we need to first prepare the media which contain all the nutrient , vitamins ,
organic and inorganic salts and the growth factors also so that the plant can easily grow in our
laboratory. There are different type of medias are used in plant tissue culture like MS media, N6
media, B5 media and B2 media. Here we are making the MS- B2 media.
Requirements:
Chemical requirements:
All the stock solutions (stock A to stock E)
Autoclaved Distilled water
Agar powder
Sucrose
Hormone (BAP)
Procedure:
1. Test tubes and beakers were taken and washed with the labeolin.
2. Then the test tubes and beakers were dried and cotton plugs were prepared for the test
tubes.
3. Then the MS- B2 media was prepared according to the chat and the ph of media was
maintained up to 5.7.
4. Then the prepared media was distributed equally among the test tubes and the test tubes
are plugged in with the help of cotton plug.
5. All the tubes were autoclave for 25mint.
6. Then all prepared media was placed in the cultured room for further use.
Fig: after the media was prepared and was stored for further use in the culture room.
Conclusion: The MS- B2 media is prepared and stored for further use.
Expt: 2
Aim of the experiment:
Initiation of Rauvolfia serpentina.
Requirements
Chemical requirements:
Bavistin
Hgcl2
Autoclaved distilled water
Procedure:
1. The explant was taken from the in vivo condition and soaked in 0.1% of Bavistin
solution for 10 to 15 minutes.
2. After this the explants were washed with running tap water for removal of excess
Bavistin solution.
3. Then the explants were washed with the autoclaved distilled water inside the laminar
air flow and transfer the explants were transfer into the UV sterilize beaker.
4. Then the explants were surface sterilized with the Hgcl2 for 2 to 4 minutes.
5. Then the explants were washed with autoclaved water for 3 times.
6. The excess part of the explants was cut with the sterilized forceps and blade.
7. Then it was inoculated inside the test tube containing media.
Fig: initiation of Rauvolfia serpentina
Expt3
Requirements:
Given sample
Petri plates
Forceps
Blade
Autoclaved tissue papers
Culture tube containing media
Gas burner
70% ethanol
Principle:
Due to the depletion of nutrients present in media we have to transfer the plant into another fresh
media for better growth and development. Then the plant is again transfer into the rooting media
for root development.
Procedure:
1. The sugar cane bunch was taken out from the old media and placed on the tissue paper.
2. Then the bunch was cleaned with the help of forcep and blade.
3. The bunch was taken by the help of forcep and was placed into the culture tube
containing fresh media.
Conclusion:
The plant is placed in the culture room after transferring into the fresh media for further growth.
Requirements:
Leaves of Ocimum gratissimum
Chopping board
Knife
Weighing balance
Tray
Clevenger apparatus
Distilled water
Principle:
Essential oils are the pure and volatile extraction from plant. It can be extracted from the seeds,
flowers, leaves, fruits and root also. They are having low boiling temperature and volatile at
room temperature. We can use this oil in cosmetics and also in pharmaceutical industries. We
can extract the oil by the help of Clevenger apparatus. This apparatus is also known as
hydrodisstilation apparatus because in this we boil our sample with the water to collect the
essential oil. The principle of Clevenger apparatus is on the basis of the method commonly used
in obtaining the volatile oil from plant product. This is carried out either by passing steam or
water through a suitable vessel containing the plant material and condensing the steam or by
boiling the material with water in a suitable vessel distilling and collecting the distillate. The
volatile oil is carried over with the steam and condenses with it. Being only slightly soluble in
water it subsequently separates from the aqueous portion of the distillate in layers.
Procedure:
1. 75gm of leaf samples were taken and finely chopped with the help of knife.
2. Then the Clevenger apparatus was washed properly and set accordingly.
3. The finely chopped samples were taken and placed inside the apparatus with some
amount of water.
4. Sample was boiled and the oil was collected.
Fig1: Clevenger apparatus
Observation:
400µl of essential oil is collected.
Calculation:
Conclusion:
400µl of essential oil is collected from 75gm of given sample and the yield is 0.533%.
Expt: 5
Requirements:
Chemicals requirement:
Methanol
Principle:
For non volatile secondary metabolites, we use an apparatus know as soxhlet apparatus.
Secondary metabolites are extracted semi continuously with organic solvent like methanol,
acetone, hexane etc. solvent is heated and volatized then is condensed above the sample. Solvent
drips onto the sample and soaks it to extract the secondary metabolite constituents at 15-20 min
interval the solvent is siphoned to the heating flask to start the process again.
Procedure:
1. 100gm of given sample was taken and chopped with the help of knife.
2. Then the chopped sample was put inside the apparatus along with the solvent (methanol)
and distillation unit was fit above the apparatus.
3. Then the whole system was switched on and running for next 72 hours. Then the extract
was collected and filtered. Again the reflux was set and run for two complete siphon and
the reflux methanol was collected.
4. The extract was dried and measure the amount of content.
Fig1: Soxhlet apparatus, fig2 and fig3: extract collected from Curcuma
amada
Observation:
Wt. of blank beaker = 51.99gm
Calculation:
Amount of extract from 10gm of given sample = 52.74- 51.99 = 0.75gm
Conclusion:
Requirements:
Bacterial culture
Nutrient media
Distilled water
Conical flask
Cotton
Gas burner
Petri plate
Weighing balance
Procedure:
1. 1.3gm of nutrient broth powder and 1.5gm of agar powder was weighed. Then mixed
with 100ml of distilled water to make the media and the media was autoclaved for
15minutes.
2. Then the media was poured into the petriplate and leave it for solidification.
3. The bacterial culture was spread on the media containing petriplate and the well was
made by the well maker.
4. Then the essential oil was poured into the well and the plate was stored for incubation.
Observation:
Conclusion:
From this above experiment we can know the antimicrobial activity of C. amada essential oil.
Expt: 7
Requirements:
Container
Bricks
Sand
Green leaves and dry leaves
Paper wastes
Cardboards
Water
Fresh cow dung
Straws
Gunny bag
Procedure:
The brick pieces were placed on bottom of the container, and then the sand was
placed on the top of the brick. Then the layer was prepared by putting the cow
dung, paper waste, green leaves, straws and dry leaves respectively. Then this was
covered by the gunny bag and was stored for few months.
Bioinformatics:
Bioinformatics is a inter disciplinary field of science in which we can study the biological
problem by the help of the information technology, he computer science, mathematics and the
statistic.
It is a field that holds great potential for revolutionizing biological research in the coming
decades. Development of tools for elucidation of the function and interaction of all genes
products in cell by integration of disparate fields of biological knowledge and a variety of
complex mathematical and statistical tool and techniques.
Database:
A structured collection of data held in computer storage especially one that incorporates software
to make it accessible in a variety of ways; transfer any large collection of information.
1. Primary database
2. Secondary database
3. Composite database
Primary database:
Secondary database:
COGS and CluSTRr for protein
FlyBase for genome sequence
CATH and SCOP for protein structure
Composite database:
NCBI
OWL
nr
1. Dynamic programming
2. Heuristic method: that is able to produce an acceptable solution to the problem. Example-
BLAST
3. Probabilistic method
Scoring of alignment:
For DNA- DNA identify matrix
Phylogenetic Tree:
We can analysis the Phylogenetic of the sequences by different method
(Q)How to do this?
Ans: uniprot
Sequence
Open (phylogeny)
Tree