ENG 10 LECTURE SECTIONS C AND T
PROF. MABINI DG. DIZON
(LECTURER)
SAMPLE Materials and Methods in Listed Form
The Effect of Nami (Dioscorea hispida Dennst.)
Crude Extract on the Internal Organs
of Tadpoles
MATERIALS AND METHODS
A. Materials
nami (D. hispida) extract
48 tadpoles
8 wide-mouthed bottles
anhydrous sodium sulfate
syringe with needle
razor blade
fume hood
mortar and pestle
chloroform
2% KOH
ethanol series (95% and 100%)
10% formalin
glycerol series (20%, 50%, and 80% and 100%)
Bouin’s solution
alizarin red stain
dissecting microscope
camera with film
(Note: Materials appear in listed form especially for a
research proposal. This is so since it is a lot easier or more systematic to
gather first all the materials needed before performing the experiment.
A disadvantage if materials are combined with the procedures is that,
just filtering through the paragraphs to identify the materials can be
another hard task. In descriptive studies, however, materials and
methods are combined.)
B. Methods
1. Preparation of plant Extract
a. Tubers of Diocorea hispida will be collected in
Batangas, cleaned, thinly sliced and air-dried
for three days.
b. The tubers will then be air-dried in an oven set
at 37 – 40 0C for another three days or until it is
crisp enough to be ground to a fine powder
form.
c. The powdered material will be soaked in
chloroform for three days with occasional
stirring.
d. The filtrate obtained will be passed through an
anhydrous sodium sulfate.
e. The resulting thick oil extract will be air dried
under a fume hood and used as the test
material.
2. Test Animals
a. A total of 48 tadpoles will be obtained from the
greenhouse area of the Biological Sciences
building.
b. These tadpoles will be placed in wide-mouthed
bottles.
3. Experimental Procedures
a. The tadpoles will be divided into four treatment
groups, namely:
legless tadpoles;
tadpoles with small hindlegs;
tadpoles with larger hindlegs; and
tadpoles with both the hindlegs and the
forelegs.
b. For each of the treatment groups, 6 tadpoles
will be considered and placed in separate
bottles with labels.
c. Another set of tadpoles will be injected with the
nami extract (about 1 to 2 milliliters) everyday
for two days.
d. Fixation of Tadpoles for Skeletal Examination
d.1. Three tadpoles from each treatment
group, including the control, will be
dehydrated in 95% ethanol for four
days; followed by another four days in
100% ethanol.
d.2. The tadpoles will be eviscerated prior to
clearing in 2% KOH for 24-35 hours.
d.3. After clearing, the tadpoles will be
stained with alizarin red for three to four
hours or until the bones become more
vivid than the surrounding tissues.
d.4. The tadpoles will be drained, destained
and dehydrated in the series of 20%,
50% and 80% glycerol for 24-28 hours in
each concentration and 18 hours in
100% glycerol.
d.5. The tadpoles will then be subjected to
skeletal examination and
photographed.
e. Fixation of Tadpoles for Soft Tissue Examination.
e.1. The tadpoles for soft tissue examination
will be fixed in Bouin’s solution for one
week and hand-sectioned using a razor
blade.
e.2. The sections will be placed in 10%
formalin, examined under a dissecting
microscope, and photographed.
4. Analysis of Data
a. Pictures of the animals treated with nami
extract will be compared to those of the
control.
b. Any abnormality or deviation from the normal
features like the malformation of a bone
structure will be noted and considered as
significant effect of the test material.
C. Detailed Schedule of Activities
Date Activity
April 8, 2006 Procurement of tubers
April 9 Preparation of tubers
April 16 Procurement and sorting of
tadpoles
April 18 – 19 Testing of the extract on
tadpoles
April 22 Dehydration of tadpoles in
95% ethanol, fixation of
tadpoles in Bouin’s solution
April 26 Dehydration in 100% ethanol
April 29 Rinsing of tadpoles in water
and hand-sectioning;
placement of sections in 10%
formalin
April 30 Clearing of tadpoles in 2%
KOH and draining in 20%
glycerol
May 2 Destaining in 50% glycerol,
soft tissue examination
May 5 Dehydration in 80% glycerol
May 7 Further dehydration in 100%
glycerol
May 9 Skeletal examination
May 13 Organization/analysis of
data
May 25 Writing of the first draft
June 5 Submission of the first draft
(Note: The Methods can consist of the main procedures
relevant to the experiment. If the procedures are long, sub
classification is encouraged. The Methods also contain the manner by
which data will be analyzed, e.g. statistical tool, as well as the Detailed
Schedule of Activities. Note that statements are in the passive voice,
future tense.
The Procedures are chronologically numbered or
labeled.
The Detailed Schedule of Activities, meanwhile,
affords the researcher to monitor if he/she is on schedule, ahead of
schedule or lagging behind.)
(Prepared by MDG Dizon, 81810)