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Section: Plant Lab Protocols
 
     
 
Plant Lab Protocols
 
   
 
Methodology for Plant Science

Science grows through the nurturing with appropriate methodologies. Choice of experimental methods in research is important to yield satisfactory results.
 
Subjects like physiology, genetics, pathology, microbiology are now getting expressed in a common language of chemistry, and in common procedures. Quite a number of books are available on general and clinical biochemistry and also on specialized topics such as proteins, nucleic acids, etc. Although there are a number of biochemical procedures followed in plant science research, details are found scattered in various journals and in some science works.
 
There is obviously a need for a source which in one place provides a various biochemical procedures that are in frequent use in agricultural research.

The outcome of our endeavour is the ePlantScience.com Plant Lab Protocol covering protocol useful for scientist working in area of plant sciences.

Although we have taken enough care to include operational details, it is necessary for the individual scientist to get conversant with the methods properly.

ePlantScience is not respondsible for any kind of inappropriate use of the methodologies given on this website.

   
  Carbohydrates  
   
Ä Color reactions of carbohydrates
Ä Determination of amylose
Ä Determination of fructose and inulin
Ä Determination of glucose by glucose oxidase method
Ä Determination of reducing sugars by Nelson-Somogyi method
Ä Determination of total carbohydrates by anthrone method
Ä Estimation of cellulose
Ä Estimation of crude fiber
Ä Estimation of hemicelluloses
Ä Estimation of pectic substances by:
Ä Colorimetric method
Ä Gravimetric method
Ä Estimation of reducing sugar by dinitrosalicylic acid method
Ä Estimation of starch by anthrone reagent
Ä Phenol suplhuric acid method for total carbohydrate
 
 
 
   
  Lipids  
    Ä Determination of iodine value of oil
Ä Determination of peroxide value
Ä Estimation of free fatty acids
Ä Estimation of oil in oilseeds
Ä Estimation of volatile acids
Ä Identification of quantification of fatty acids
Ä Saponification value
 
 
 
   
  Amino Acids & Proteins  
    Ä Ammonium sulphate fractionation of proteins
Ä Determination of net protein utilization, digestibility and biological value
Ä Electrophoresis of proteins in starch gels
Ä Enzyme-linked immunosorbant assay (ELISA)
Ä Estimation of lysine
Ä Estimation of methionine in food grains
Ä Estimation of proline
Ä Estimation of total free amino acids
Ä Estimation of tryptophan
Ä Fluorography of polyacrylamide gels
Ä Identification of proteins
Ä Immunodiffusion in agrose gels
Ä In vitro translation assay
Ä Isoelectric focusing
Ä Mass Screening test for high-lysine grains
Ä Nitrogen analysis by micro-Kjeldahl method
Ä Polyacrylamide disc gel electrophoresis
Ä Polyacrylamide-sodium dodecyl sulphate slab gel electrophoresis (SDS-PAGE) of proteins
Ä Preparation S-30 extract for protein synthesis in vitro
Ä Production of antiserum
Ä Protein (Western) blotting
Ä Protein digestibility in vitro
Ä Protein efficiency and net protein ratio
Ä Protein estimation – Bradford method
Ä Protein estimation by Lowry’s method
Ä Quantification of proteins in polyacrylamide gels
Ä Rocket immunoelectrophoresis
Ä Sample preparation for amino acids estimation
Ä Silver staining of proteins
 
 
 
   
  Enzymes  
    Ä Acetyl cholinesterase
Ä Aminotransferases
   i) Asparatate amino transferase
   ii) Alanine aminotransferase
Ä Amylases
Ä Ascorbic acid oxidase
Ä Asparagine synthetase
Ä Catalase
Ä Cellulases (C1 and Cx)
   i)   Viscometric method
   ii)  Colorimetric method
   iii) Combined assay
Ä Glutamate dehydrogenase
Ä Glutamate synthase
Ä Glutamine synthetase
Ä Indole acetic acid oxidase
Ä Isoenzyme analysis
   i)   Esterase
   ii)  Polyphenol oxidase (PPO)
   iii) Peroxidase
   iv) Acid phosphatase
   v) Glutamine synthetase
   vi) Phosphoenol pyruvate carboxylase (PEP Case)
   vii) Glutamate dehydrogenase
   viii) Indolylacetic acid oxidase
   ix) Malate dehydrogenase
   x) Sucrose synthase
   xi) Aspartate aminotransferase
   xii) Glutamate synthase
Ä Lipase
Ä Malate dehydrogenase
Ä Nitrate reductase
Ä Nitrite reductase
Ä Papain
Ä Pectin methyl esterase
Ä Peroxidase
Ä Phenylalanine ammonia lyase
Ä Phosphatases
   i)  Acid phosphatase
   ii) Assay using glycerophosphate
Ä Phosphoenol pyruvate carboxylase
Ä Polygalacturonase activity
Ä Polyphenol oxidase
Ä Ribulose biphosphate carboxylase
Ä Sucrose synthase
 
 
 
   
  Nucleic Acids  
   
Ä Cultivation of lambda (l) phage
Ä DNA electrophoresis in agarose gel
Ä Estimation of DNA
Ä Estimation of RNA
Ä Extraction of lambda (l) DNA
Ä Extraction of plant DNA
Ä Extraction of plant total DNA (Rapid method)
Ä Isolation of deoxyribonucleic acid (DNA)
Ä Isolation of messenger RNA by affinity chromatography
Ä Isolation of plasmids
Ä Isolation of ribonucleic acid
Ä Rapid isolation of plasmid DNA (Miniscreen)
Ä Recovery of DNA fragments from agarose gels
Ä Restriction of DNA
Ä Southern blotting
 
 
 
   
  Vitamins  
    Ä Ascorbic acid
   i)  Volumetric method
   ii) Colorimetric analysis
Ä Niacin
Ä Riboflavin
Ä Thiamine
 
 
 
   
  Pigments  
    Ä Carotenes
Ä Chlorophylls
Ä Curcumin
Ä Lycopene
 
 
 
   
  Phenolics  
    Ä Capsaicin
   i)  Colorimetric Method
   ii) Spectrophotometric Method
Ä Chlorogenic acid
Ä Lignins
Ä Phenols
Ä Tannins
   i)  Folin-Denis Method
   ii) Vanillin Hydrochloride Method
 
 
 
   
  Antinutritional Factors  
    Ä Cyanogens
Ä Gossypol
Ä Phytic acid
Ä Trypsin inhibitor
 
 
 
   
  Plant Hormones  
    Ä Ethylene
Ä Indoleacetic acid
 
 
 
   
  Nitrogen Fixation  
    Ä Allantoin and Allantoic acid
Ä Hydrogen evolution
Ä Leghaemoglobin
Ä Nitrogenase
 
 
 
   
  Separation Procedures  
    Ä Analysis of aflatoxins
Ä Column chromatography
Ä Paper Chromatography
Ä Thin layer chromatography
 
 
 
   
  Cell Fractionation  
    Ä Fractionation of cellular components
Ä Isolation of chloroplasts
Ä Isolation of mitochondria
 
 
 
     
 
     
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