Estimation of Protein from Germinated and Non Germinated Seeds-Modern Biotechnology and Applications-Project Report, Study Guides, Projects, Research of Biotechnology

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PROJECT REPORT ONA

In the Partial fulfillment of B.Sc. (Biotechnology)^ “ Estimation of Protein from Germinated and^ Non Germinated Seeds”

I wish to express my gratitude towards all those who have been helpful to me in getting this mightily task of project done up. I would like to acknowledge the overweening support, from both teacher ACKNOWLEDGMENT

and student alike, enjoyed by biochemistry this gave me the confidence & courage to effect on the project work. I would like to express provided by Miss Ritu Mittal and Miss Deepka Verma from their valuable help guidance during the course of project work. my appreciation for the generous support

Parul Tyagi

    IntroductionInstrument and RequirementPreparation of ChemicalsPrinciple of Colorimeter INDEX

      Source of Protein UsedProcedure for Estimation of proteinTable of Non Germinating SeedTable of Germinating SeedGraph of Non-GerminatingGraph of Germinating

  ResultBibliography

Protein is primary a building block for body our needs go down once we have reached adulthood and have stopped growing. At that time protein is required for maintenance and repair. INTRODUCTION

protein different from human protein through both are made of amino acid. High protein diet are especially toxing to the liver and kidney. When people eat too much protein. They take in more nitrogen than they need.^ The actual fact is that protein of each animal is unique. Animal

minerals amino acid and carbohydrate. varities red clours, radish, mung beans peas.^ Seeds are a good source of protein essentially fatty acid, vitamin E, You can sprout seed bean grain, nuts, some of most popular Everyone is encouraged to eat more eggs because of The proteins

they contain. Remember that once egg cooked their protein coagulate, making then less readily available to the body. There are also several other health problems associated with consumption of eggs.

Colourimeter is the electromagnetic radiation in visible reason of spectrum. Colorimeter can help find the concentration of substance. Since the amount and colour of light that is PRINCIPLE OF COLORIMETER measurement of the wavelength and the intensity of absorb or transmitted depend on the property of solution. solution with the amount that can get through a sample of pure solvent. A colourimeter contain photocell able to detect the amount of light which passes through the solution under investigation A colourimeter is an instrument that compares the amount of light through a PROCEDURE: 1. 2. 3. 4. Switch on power supply to colorimeter for 10 min.Select the desired filter.Adjust 0 before calibration of InstrumentFirst adjust 0 with the help of distilled water by placing it in holder and by adjusting 100% transmission.

5. 6. 7. Switch off shutter and remove the cuvette containing distilled water.Now adjust 0 with the help of black selution by placing it in holder and adjusting transmission.Switch off shutter and remove the cuvette containing blank solution.

1. 2. 3. ELECTRIC BALANCEREFRIGRATERCOLORIMETER INSTRUMENT

4. MANUAL BALANCE REQUIREMENT

9. BSA

 1. 2. ALKALINE NaTaking NaoH (200mg NaoH in 100ml distilled water)Dissolve 2gm Na 2 Co^ PREPARATION OF CHEMICALS 2 Co (^33) in NaoH.   1. 1. 2. CuSo1 gm pot tartrate in 100 ml500 mg CusoALKALINE Cuso1 ml Cuso 4 REAGENT 4 + 50 ml alkaline Na 4 dissolve in 1gm potassium tartrate. (^42) Co 3

  MIXTURE SOLUTION 15 mg in 3 ml 0.1 N NaoH SolutionFOLIN’S REAGENT 0.5 ml folin reagent make dilute by adding 0.5 ml distilled water.

A) PREPARATION GERMINATING & NON GERMINATINGESTIMATION OF PROTEIN BY “LOWRY OF^ METHOD” STANDARD SOLUTION FOR

    Firstly taking germinating and non germinating seedsThan crush the seed by pastle-mortar and prepare the reagentCrushed seed mixed in 0.1N NaoH solution and then prepare Standard solution.Taking 6 test tube and mark them o, 0.2, 0.4, 0.6, 0.8 or 1 ml and

  add standard solution according to test tube in which conc. is 1mg,^ 2mg, 3mg, 4mg & 5mg.Then add to it 0.1 N NaoH in each test tube according to 1, 0.8, 0.6, 0.4, 0.2, 0 ml in each test tube.Then add 3 ml freshly prepared alkaline CuSo 4 reagent in each test

 tube and remain constant for 10 min.After 10 min add 0.5ml folin’s reagent and take final volume 4.5ml then stand it for half an hour.

  After Stand is for half an hour, a purple colour variation takes place in different test tube.Then note its optical density.

8. 9. 10. Rinse the cuvette with the help of distilled waterPet standard solution in cuvette holder and place it in holder and then ineasured reading on the calorimeter as oppical reading.Switch off shutter & remove the cuvette

11. Take the reading and plot the graph.

S.No.^ GRAM NON GERMINATING SEED Mix Non Sol TABLE FOR NON GERMINATING SEED of 123 Germinating^ seed (^0) 0.20.4 0.1^ NaoH^ Sol (^1) 0.80.6 Alk.^ Phosphate^ reagent3 ml3 ml3 ml Incubation^ period10min10min10min Folin^ reagent0.50.50.5 0 Total^ Const.1mg2mg Incubation^ period30min30min30min Optical^ Density0.190.26____ 456 0.60.81ml 0.40.2 0 3 ml3 ml3 ml 10min10min10min 0.50.50.5 3mg4mg5mg 30min30min30min 0.340.450.

S.No. Mix Non Germinating Sol of LOBIA NON GERMINATING SEED 1234 seed (^0) 0.20.40.6 0.1^ NaoH^ Sol (^1) 0.80.60.4 Alk.^ Phosphate^ reagent3 ml3 ml3 ml3 ml Incubation^ period10min10min10min10min Folin^ reagent0.50.50.50.5 0 Total^ Const.1mg2mg3mg Incubation^ period30min30min30min30min Optical^ Density0.300.410.54____ 56 0.81ml 0.2 0 3 ml3 ml 10min10min 0.50.5 4mg5mg 30min30min 0.630.

S.No.^ GRAM GERMINATING SEED Mix Non Sol of TABLE FOR GERMINATING SEED 123 Germinating^ seed (^0) 0.20.4 0.1^ NaoH^ Sol (^1) 0.80.6 Alk.^ Phosphate^ reagent3 ml3 ml3 ml Incubation^ period10min10min10min Folin^ reagent0.50.50.5 Total^ Const. (^0) 1mg2mg Incubation^ period30min30min30min Optical^ Density0.400.47____ 456 0.60.81ml 0.40.2 0 3 ml3 ml3 ml 10min10min10min 0.50.50.5 3mg4mg5mg 30min30min30min 0.540.620.