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					Partial Characteriztion of the biodegrading ability of the fungi Xylaria
sp. and its four mutants on natural rubber, chicken feathers and
polystyrene.

Objectives:
  1. to    determine  if  Xylaria   sp    mutants    and   wildtype        can
      degrade/consume/ assimilate natural rubber as a carbon source
  2. to    determine  if  Xylaria   sp    mutants    and   wildtype        can
      degrade/consume/ assimilate chicken feathers as a carbon             and
      nitrogen source
  3. to    determine  if  Xylaria   sp    mutants    and   wildtype        can
      degrade/consume/ assimilate polyurethane as a carbon source
  4. to compare the degradation capability of the wild type and            the
      mutants.


Outline of methodology

  I.       Preparation of Inoculum
              Isolate the Xylaria sp. by culturing it in a Potato Dextrose Agar
        (PDA) medium. Adjust to pH 5 and incubate at 25˚C. After 2-3 days,
        transfer the fungi into test media.

  II.     Preparation of Pollutants
         A. Polystyrene
                  - Cut 1x1 cm squares from plastic cups.
         B. Chicken feather
                  - Obtain fresh feathers from Gallus gallus sp. Wash and
                     sterilize in an autoclave.
         C. Natural Rubber
                  - Obtain a unused rubber latex glove. Cut 1x1 cm of the
                     gloves.

  III.    Preparation of Test Media*
         A. For plates:
                    Prepare 2 sets of plates containing 20 ml PDA in triplicate
             (6 plates per mutant/wild type, per pollutant tested). Add 0.5%
             glucose in set A and add 0.5% of the liquid pollutant in set B.
             Adjust to pH 5 by adding small amounts of either 0.1M NaOH or
             0.1M HCl. Inoculate the fungi using agar discs from the PDA
             plate described in I. Agar discs will be obtained by cutting a 2-3
             day-old inoculum on the peripheral area of a colony using a 5-8
            mm diameter cork borer. The agar discs will be transferred to
            the PDA plates of sets A and B by using a sterile toothpick.
            Incubate at 25˚C.
                  Observe the colony growth in set B and compare it always
            with set A. Set A will be the control group and it would indicate
            whether the fungi transferred is active. Then measure the colony
            growth by its diameter.

        B. For flasks and test tubes:
                   Prepare 2 sets of flasks containing 50 ml Mineral Medium
            each, in triplicate. Add 0.5% glucose in set A and B. Adjust to pH
            5 by adding small amounts of either 0.1M NaOH or 0.1M HCl.
            Then add the solid pollutant in set B only. Inoculate the fungi by
            using ………….. When all the glucose has been used up and the
            fungi had grown into a considerable mass as examined visually,
            add another MM + 0.5% glucose in set A only, leaving the set B
            flasks to utilize the solid pollutants as the sole carbon source.
            The extent of colonization should be carefully examined every
            day until rate of colony growth can be predicted (growth in
            mm/day). But if otherwise, continue adding the MMG (mineral
            medium+0.5%glucose) to both sets of flask until the fungi has
            grown and thrived. Incubate for 50-80days, with the flasks in a
            room with more or less 250C in temperature, under shaking
            conditions, while the test tubes in an incubator with 25 0C in
            temperature as well.

  IV.    Remove solid pollutants from the culture medium and examine
         under a scanning electron microscope (SEM).


Note:
* - this step is intended for each mutant and for the wild type. Since we
have 4 mutant strains and a wild type, this step will be repeated five times
multiplied with the number of pollutants to be used.

				
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posted:6/27/2011
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Mary Bernadette Vallesfin Egloso Mary Bernadette Vallesfin Egloso English Teacher http://adelaide17madette.multiply.com/
About My friends call me Addie. I want to become a doctor someday and serve my countrymen after studying medicine in the Philippines. I also want to become a sophisticated investor and business owner someday. I truly believe in what Robert Kiyosaki said in his books. It is very important to keep on improving oneself, as we live in this dynamic and competitive world. I love swimming and singing.