Jatropha

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INTRODUCTION: - Agriculture is the backbone of our country. If we potentially utilize our waste land in the proper and scientific way; we have to solve our poverty level and unemployed generation. For a better world to live in, we need pollution free air, safe water, nutritional food, healthy environment and greenery around us. Without sustainable and secure livelihoods will be feeding on each other leading to poverty, pollution, poor health, biodiversity, erosion and land degradation.

Energy and greenery environment is an outlook for everyone’s not only for the development but for the security and safe guard of existence. Presently our main resource has also limited available storage. If we do not take proper step in time, we shall face lots of problems.

After a continued study and research work at Purulia district, it has been found that powerful alternative source of bio energy that will be helpful in environment remedies as well as it will be pathfinder in our future.

The district of Purulia which has abundance of waste land more than laces of acre and its surrounding needs such type of farming related with Jatropha. And it is observed that because cultivation of jatropha, the erosion of waste land has substaintially been reduced. Hence cultivation in wasteland will automatically help in soil conservation and will create lot of employment opportunity in rural areas.

Jatropha is a draught resistant; photo- insensitive perennial shrub belonging to the family Euphorbiaceae is attracting increased attention as source of bio-diesel. This is a non-grazing hardy plant can be grown in wasteland. It is planted in the month of June -sept. during on set of monsoon. The seed contain oil which is used as a bio-diesel, 25-30% oil is present and its oil cake is not used as fodder because due to presence of toxic element. It is used for nitrogen enriching organic manure.

Jatropha curcas L (Euphorbiaceae) is a large shrub or tree native found & utilized throughout most of the tropical and subtropical region of the world. Jatropha curcas is the most important amongst tree borne oilseed viz karanja, neem, jojoba, etc having potential growth in wasteland. Morever, Jatropha exhibit a wide environmental tolerance; it is found in both seasonally dry tropic as well as equatorial region, and is well adopted for cultivation within the vast area of marginal and degraded lands in the semi arid and dry tropic.(At Purulia and Bankura W.B) at our company Jatropha project exhibit high potentially grow in this dry zone.

India has nearly 60 million hectares of waste lands, of which 30 million hectares can be made available for Jatropha plantation capable of yielding 60 million tones of bio-diesel annually on a sustained basis. Bio-diesel is a carbon neutral and also has other environmental advantages as under:-

* Particular matter (PM) emission are30% lower than fossil diesel.
* Carbon dioxide emission are 78% lesser than fossil diesel
* Bio diesel is a clean and renewable fuel
* Carbon monoxide exhaust emissions are 50%lower than diesel.
* Hydrocarbon exhaust emissions are 93% lower than diesel.
* The overall ozone (smog) forming hydrocarbon emission are 50% lesser than that of bio diesel.
* Bio-diesel reduces the health risks associated with diesel by releasing decreased level of ploy cyclic aromatic hydrocarbons (PAH) and nitrate PAH compound by 75 to80.

CARBON SEQUESTRATION CDM BENEFITS

In the 1997 World Earth Summit held at Kyoto, Japan 160 countries of the world had participated. The motive was to negotiate binding limitations on greenhouse gases for the developed nations pursuant to the objective of the Framework Convention on Climate Change of 1992. The outcome was the Kyoto Protocol, in which the developed nations agreed to limit their greenhouse gas emissions, relative to the levels emitted in 1990 or pay a price to those that do. At this point comes the CARBON TRADING. The idea was to make developed countries pay for their wild ways with emissions while at the same time monetarily rewarding countries with good behaviour in this regard. Since developing countries can start with clean technologies, they will be rewarded by those stuck with 'dirty' ones. This system poises to become a big machine for partially transferring wealth from wealthy, industrialized countries to poor, undeveloped countries.

Trading in carbon credits is an innovative way to keep carbon dioxide emission from industries, which leads to global warming-in check.

This allows industries in developed countries to off-set their emissions of carbon dioxide by investing in reforestation and clean energy projects in developing countries such as large-scale tree plantations as a lucrative alternatives to reducing emissions or acquire such Carbon Credit from companies in India doing such Business. This is what makes trading in carbon credits such a great business opportunity. Foreign companies which cannot fulfill the protocol norms can buy the surplus credit from companies in other countries. This lead to a flourishing trade called CARBON TRADING. India is considered as the largest beneficiary, claiming about 31 percent of the total world carbon trade through the Clean Development Mechanism (CDM). It is expected to rake in at least $5 billion to $10 billion (Rs.22,500 crore to Rs.45000 crore) over a period of time.
How is it Beneficial for Sunplant
Planting 100,000 hectares of new forest can remove one million tones of carbon per year from the atmosphere. As trees grow, they adsorb carbon dioxide from the air. We at SUNPLANT have been planting trees and as such we are reducing the carbon dioxide emission in the atmosphere. Moreover we are also venturing into BIO-DIESEL & ORGANIC FERTILIZERS, which is another environment friendly activity. Since we are working towards DECARBONIZATION, trading in Crabon Credits by us would be huge.

We at SUNPLANT have been using new and modern concepts ad technology to achieve our MOTTO & VISSION OF A CLEAN AND GREEN FUTURE.

Key advantages of the process of CO2 sequestration using algae

* Owing to the fact that high purity CO2 gas is not required for algae cultivation, flue gas containing CO2 and water can be fed directly to the photobioreactor.
* Power plants that are powered by natural gas or syngas have virtually no SO2 in the flue gas. The other polluting products such as NOx can be effectively used as nutrients for micro algae.
* Micro algae culturing yields high value commercial products that will offset the capital and the operation costs of the process. In addition to biofuels, algae are also as the starting point for high-protein animal feeds, agricultural fertilizers, biopolymers / bioplastics, glycerin and more.
* Algae can grow in temperatures ranging from below freezing to 158ºF.
* The entire process is a renewable cycle.

PROJECT ON ALGAEL BIOFUEL :
1. After technical discussion with IGE following points are considered for PROJECT - ALGAE in presence of technical
experts from IGE and SPAL( +) ;
1st phase ;

* Identification and characterization of local fresh water algae
* Growth study ; Media Selection ,Light and Ph standardization in respect to selected strain of algae
* Biomass ; Lipid and Protein content (yield of selected strain)
* DNA profile of selected strain
* Long term storing ( deep freezer)

Duration ; 3-6 months
2nd Phase ; Pilot Project ( Micro Algal Biotechnology )

* Bioreactor for micro algal photosynthesis and Cultivation system study
Aim ; Designing of Bioreactor for optimum growth and Lipid production
* Open pond system ; Micro algal Growth and Cultivation system study
Aim ; Optimum growth and Lipid prodction

Duration ; 12 months

PROJECT REPORT
1. Following technical information described on “Project on Algael Biofuel” as per project plan mentioned above for
evaluation of said projct launched on 01-03-2010 at IGE
First phase of the project completed as follows ( by 29-03-2010) ;

1. Four local fresh water algael strain was isolated and characterized.
2. Growth studies were done .
3. Biomass and Oil extraction are going on.
4. Open pond observation of algael growth ( all four strain together ) for Biomass yield are going on ;

Four local fresh water algael strain was isolated and characterized are as follows ;

1) Selected Strain- Chlorella sp.

INTRODUCTION

* Chlorella microalgae is a tried and tested oxidation method for the treatment of effluent.
* Chlorella contains between 14 and 29% oil.
* Chlorella unicellular and grows prolifically doubling its biomass weight in few hours of sunlight.
* Chlorella vulgaris is known as one of the fastest growing microalgae and includes a reasonable amount (14-30%) of triacylglycerol for biodiesel production.
* High carbon fixing capacity per unit dry weight biomass.
* Resist & suppress colonization of potentially invasive species.
* Co-products, nutrient recycles, wastewater treatment -used in the nutrient removal from wastewaters

Collection and identification

The collection of water samples begun from 10th Feb to 12th Feb 2010.
Corresponding screening of species and confirmation was done between
11th Feb to 19th Feb 2010.

Water samples are collected from near by ponds of our college locality, Badu,Madhyamgram. The water samples were screened for identification of species and hence chlorella was identified and was further screened for confirmation.

1. Isolation
19th Feb 2010, Soil separation from algal mass was done.
Treated with Tween- 20 and salinity treatment was done to remove soil and protozoa. Specific media like Bristol was prepared for isolation of species. Serial dilution was performed for better isolation of species. Streaking inoculated species to specific media was done and kept for incubation in the environment.After 4 days growth was observed in the petriplates, dated More samples are collected from the nearby area. Serial dilution, salinity treatment was done in same way.
Purification
Serial dilution was done. Specific media Bold Basal media was made on and plates were made. Species were inoculated in liquid culture of both Bristol and Bold Basal media. growth was maintained in the jam bottles. This was done with proper media and providing proper sunlight intensity by maintaining light interval Further purification was done by treatment with salts like sodium carbonate and sodium bicarbonate on Proper light stress was provided and grow was observed and huge pure culture was observed,

Biomass cultivation

1 litre media was prepared and all necessary apparatus was arranged for
transfer for culture to the aquarium. media was transferred to aquarium, pure chlorella was inoculated, artificial light source is provided, and aeration is provided for CO2 capture, pH maintainance.The culture is maintained by proper monitoring the system.

2)SELECTED STRAIN:- EUGLENA sp.

Raw sample was collected from nearby ponds,drains,Soil etc.each sample was observed under microscope & various strains were identified.
Samples were kept in bottles in water,kept under Sunlight for the growth
observation.the samples were mainly mixed culture.euglena was identified.

Features:-

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Single Celled
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No cell wall
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Autotroph
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Motile
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Asexual reproduction
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Kingdom Protista
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Autotroph– Euglena have chloroplasts that undergo photosynthesis using light energy to build sugar.
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Flagellum– The flagellum, or tail, helps the euglena move towards light and away from predators.
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Eyespot– The Euglena has a very simple eye that senses light. This way the euglena can move untill it is in a lighted area.

Further screened for isolation.first,serial dilution is done.here,1ml of raw sample containing Euglena sp. is added into 9ml of water.then,from it again 1ml is added to another test tube containing 9ml water.upto minus four & kept in light for growth.
Others strains were also kept under study.the specific media for the growth of euglena was studied.media compositions were collected in the laboratory to make the media.media is nothing but the artificial habitat of the algae where they can grow with the help of nutrient in laboratory condition.sodium acetate can help to increase the growth rate of euglena.media was composed of the following:-

KNO3- 0.5g
KH2PO4- 0.5g
MgSO4- 0.25g
NaCl- 0.1g
FeCl3- TRACE
TRYPTONE- 5g
1LTR DISTILLED WATER.

The 1ltr media was devided into 4 bottles 250ml each.they are autoclaved.then,agar was added to 3 bottles.they were poured into 11 petriplates.the 4th bottle is kept aside without agar.it was treated as broth.the 11 plates were kept for one night for the agar to be cooled & solidified.the next day,raw samples were inoculated in the plates & kept for growth growth showed yellowish,black colony.one centre in each colony was prominent.the black coliny was screened under microscope,& growth of euglena was observed.
observation.The broth was devided into 3 parts with excess nutrient & raw samples were also inoculated in it. After 3 days,growth were observed in some plates.

They were kept for further growth.
Euglena is my topic of interest.because:-

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EASILY AVAILABLE FROM FRESH WATER SURFACE.
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CAN BE CULTURED IN LABORATORY CONDITION USING SUITABLE MEDIA.
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PROPER SUNLIGHT,CO2,WATER & MAINTAINANCE OF PH(=7) IS NECESSARY.
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LIPID CONENT IS VERY HIGH,SO OIL CAN BE PRODUCED.
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PRODUCTION OF HIGH BIOMASS IS POSSIBLE AS A MONOCULTURE.

EUGLENA CAN BE USED AS...
• FOR FOOD PROBLEM- Euglena can be used for nutritional food and supplements, which is beneficial for malnourished people especially insufficient in vitamins and protein. Our goal is to provide food assistance for developing countries.
• For environmental problems- Euglena absorbs carbon dioxide to produce valuable substances. It can capture high concentrated carbon dioxide (up to 40%) of gases exhausted from plants (power plants, iron works, etc.) to reduce carbon dioxide emission.
• For space food- From its nutritional and functional aspects, Euglena is anticipated to be used for space food.
• Euglena is able to capture carbon dioxide and produce various materials which can be used for Food, Cosmetics, Feed, Fertilizer, and Fuel. Euglena can grow under an extremely high concentration of carbon dioxide, even up to 40%. Hence, Euglena can be applied to exhaust gases from power plants, iron works, etc. to reduce carbon dioxide emission.

Eutrophication by Euglena sp.

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If a body of water is polluted when euglenas are thriving in that source,Euglenas love to feed on green algae, which feed on nitrogen, a type of waste.
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The primary producers who ingest these nutrients create toxins harmful to plants and animals, as well as decreases, sometimes detrimental decreases, in dissolved oxygen.
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When these levels become hypoxic, or detrimental, plants, fish, and marine mammals do not get the oxygen they need. Therefore, biodiversity is decreased.
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In serious cases, conditions may become anaerobic, in which case Clostridium botulinum or may grow, which produce toxins deadly to
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birds and mammals. Birds and mammals also dependent on food sources in fresh or stagnant water are at risk as well.

Production of monoculture:-

After identifying the strain of euglena,it was treated for large scale production of pure monoculture.for this,media was prepared again & its PH was measured.euglena growth can be maximum at a PH of 6.2.so it was balanced.other growth factors,i.e.CO2,aeration,light intensity(equal light & dark phase) were kept in concern while growing.
Media is poured in aquarium & the purified colony was inoculated.the cell no.was uncountable.

Strain Selected: Diatom.

INTRODUCTION:

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Diatom is a microalgae. Diatom is a member of Bacillariophyceae family.
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Diatoms are very useful in agricultural field as biofertiliser.
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Production of biofuel due to its higher amount of lipid content i.e. 45%-85%.
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Diatoms after death deposits at the base of pond and ocean bed and results in the formation of Diatomaceous Earth.
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This is a good fertilizer and the enriched source of minerals so widely used.
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It can absorb nitroglycerine. Diatoms approximately fixes one quarter of global fixed carbon and one fifth of the oxygen we breathe.
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It has multiple industrial uses.

COLLECTION:

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Local pond water collected in sterilized jam bottles.

LOCATION:
o Pond outside IGE, Thakurpukur, and other local ponds located with the help of a local person around IGE.Roadside drain water was collected.

NATURAL GROWTH & NATURAL PURIFICATION:

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Kept in jam bottles and aquarium without any nutrient for natural growth.
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Frequently water sample is tested purified.
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Precipitated algal mass pipetted and grown in fresh tap water. Kept in sunlight.

NATURALLY GROWN STOCK CULTURE

IDENTIFICATION: water samples were screened under low and high power microscope to identify the possible algal samples present in the samples.

PURIFICATION:

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Water samples observed to contain protozoa, worms, soil particles etc.
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The water sample was initially washed by TWIN 20 washing.
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Distilled water added and contaminant from the upper surface was discarded.
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Sodium chloride solution of 1%,5%,10% is added to the 100ml of raw water sample serially and mixed well.
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Diatoms precipitated at the bottom and from the upper surface the contaminants were discarded.
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Biotin was added for protozoa removal.
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Diatoms purified and kept in distilled water containing silica and sodium bicarbonate.
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Diatoms were found to be mixed with other algal species.

ISOLATION:
CHU10 & Modified combo media was prepared and plated on agar and streaking was done.Serial dilution of the purified water samples and raw water sample was streaked to observe presence of contamination and isolation and further purification.
After one day mono colony of the diatoms appeared in the form of circular
brownish colony.Mixed purified culture was also added to the liquid media
for isolation and growth. Kept in sunlight and artificial light.

ISOLATION OF THE DIATOM
GROWTH PARAMETER:

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Optimum pH for optimum growth is 7.8-8.
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Silica is an important element in growth of the species.
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5% sodium bicarbonate solution with silica resulted highest growth with salinity raised by 10% sodium chloride solution.
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Growth observed in minimal silica and excess silica and without silica.

LIQUID CULTURE: F/2 media and Modified combo media is selected for the biomass growth in aquarium with the ph of 7.8 and continuous aeration.

4) ALGAL SPECIES:-Scenedesmus sp.

INTRODUCTION:-

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It is a fresh water unicellular algae.
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It belongs to the family chlorophyceae.
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It grows well in wastewater
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Proven nutrient removal capacities
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Cheap and available
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It has protein content of 20%,lipid content of 15% & carbohydrate content of 52% respectively.
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Scenedesmus, with about 100 species is widely distributed in almost all pools of standing water.
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The number of cells in a coenobium is usually 4-8 cells which are ellipsoidal to fusiform in shape.
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Each cell is uninucleate and contains a single longitudinal laminate chloroplast with one pyrenoid.
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Each cell in a colony is capable of producing a daughter colony by division.
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A single cell may produce as many as 2,4,8,16,& 32 autospores.
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Scenedesmus sp.has been reported for waste water treatment.
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It has been used in removal of excess nitrogen & phosphorus in nutrition pollution.
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This species has can grow enormously in sunlight & variant light intensity.
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It maintains a photoperiodic cycle of 14h:10h cycle

ISOLATION & CULTURE PROCEDURE:-

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Algal samples were collected from freshwater bodies.
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Collected algae were cultured preliminarily in nutrient medium in order to examine all types of algae strains in sampled water.
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Two techniques for strain selection were utilized. First, Species were selected from phycological literatures that have been demonstrated to have a high tolerance to wide temperature fluctuations and lipid rich promising.
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The second approach involved assaying local collected strains from previously stated regions as they are likely to be adapted to certain aspects of the local climate.
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From the collected samples of water,through the screening procedure under light microscopesp.was identified.

The species was then isolated from other mixed contaminants by serial dilution,where the algal cultures were simply maintained in tap water.

To increase the biomass specified nutrient media were prepared & culture was inoculated in it.

The pH of the media is maintained between 6-7

The culture after inoculation was maintained under natural sunlight for 6-7 hrs where enormous growth was observed.
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Some of the culture were mixed with agar to give a solidified medium.
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It was then poured in petri plate.
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The previous serial dilution culture was then streaked into the plate to get single colony of the desired species.
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Growth was observed in such plates after 3days.
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Then the single colony of the species was isolated from the petri plate & transferred in an aquarium for pure mono culture growth.
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In the aquarium, the photoperiod is maintained & it is illuminated by a 40w fluroscent lamp.

MEDIA PREPARATION PROCEDURE :-

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Each of the individual media was along with the trace elements was weighed.
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It was then diluted with 850ml of distilled water with each of the specified components.
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Then the total volume was made upto 1lt.
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Then the pH of the medium is adjusted.
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For streaking the media in petri plate, 1.5gm agar is added to the medium.
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Then the media is autoclaved.
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For liquid culture or broth medium, no agar is added. The media after autoclaving is cooled & culture medium is poured directly through micropipette