DISHA DELPHI PUBLIC SCHOOL

(2023-24)

Biology Lab Manual

Class XII | Practical File
Name :

Class & Section:

Roll No.:

Instructions to Follow:

1. Students are supposed to prepare Biology practical file in Biology practical manual of
any publication.
2. All the diagrams should be drawn by pencil on plane sheet. They need to be properly
labeled.
3. Students are supposed to start every experiment from new page.
4. Index needs to be properly filled.
5. You are supposed to note down the entire content in manual as provided by school.
6. Use only blue pen to write the content.
7. All the work done should be neat and clean.
8. Observation table should be on ruled sheet, with proper boundaries, using pen.
9. Write all the experiment sequentially as provided.
10. We will not accept untidy, incomplete work.
11. Cover your files with white sheet at the time of submission.
 CLASS XII BIOLOGY PRACTICALS
(2023-24)
INDEX
S.
AIM OF THE EXPERIMENT DATE SIGN
NO.
1 Prepare a temporary mount to observe pollen germination.

2 Study plant population density by quadrat Method.

3 Study plant population frequency by quadrat method.

4 Prepare a temporary mount of onion root tip to study mitosis.

5 Isolate DNA from available plant material.

SPOTTING
EXPERIMENT NO.-1 Maize Flower (Wind Pollinated)
Salvia Flower (Insect Pollinated)
Bigonia Flower (Bird Pollinated)
EXPERIMENT NO.-2 Pollen germination on stigma
EXPERIMENT NO.-3 T.S of testis
T.S of ovary
EXPERIMENT NO.-4 Meiosis in grasshopper testis through
permanent slides.
EXPERIMENT NO.-5 T.S of blastula
EXPERIMENT NO.-6 Study of Mendelian inheritance using
seeds of different colours / size of any
plant.
6
EXPERIMENT NO.-7 Rolling Tongue – Pedigree Chart
Widow’s Peak- Pedigree Chart
EXPERIMENT NO.-8 Emasculation
Bagging and tagging.
EXPERIMENT NO.-9 Ascaris
Entamoeba
Plasmodium
Ringworm
EXPERIMENT NO.-10 Lichen
EXPERIMENT NO.-11 Homologous Organs in Plants
Analogous Organs in Plants
Homologous Organs in Animals
Analogous Organs in Animals
 EXPERIMENT NO. 1
Aim: Prepare a temporary mount to observe pollen germination.

Material Required: Fresh seasonal flowers, slide, coverslip, microscope, sucrose, Boric Acid, Magnesium
sulphate, Potassium Nitrate, beakers etc.

Theory: Pollen grains are the microspores or male gametophyte which contain male gametes. Each
pollen grain consists of two wall layers i.e outer exine and inner intine. Exine is thick &
ornamented due to sporopollenin while intine is thin, made up of cellulose. Through germ pores
intine comes out as pollen tube. Each pollen grain carries a vegetative nucleus and two male
nuclei.

Procedure
1. We will prepare a nutrient solution by dissolving 10g Sucrose, 10g Boric Acid, 30 mg
Magnesium Sulphate and 20mg Potassium Nitrate in 100 ml of water.
2. We will take few drops of solution on a clean slide and dust a few pollen grains from the stamen
of a mature flower on it.
3. We will observe the slide in the microscope after 30 minutes.

Observation
In nutrient medium the pollen grain germinate. The tube cell enlarges and comes out of the
pollen grain through one of the germ pores to form a pollen tube. The tube nucleus descends to
the tip of the pollen tube. The generative cell also passes into it. It soon divides into two male
gametes. Each male gamete is lenticular to spherical in outline.
Precautions:
1. Flowers should be freshly plucked.
2. Use clean slide to observe the pollen grains.
3. There should not be any air bubble under the coverslip.
4. Soak extra solution with filter paper.
 EXPERIMENT NO. 2
Aim: Study plant population density by quadrat Method.
Material Required: Nails, thread, meter scale, hammer, paper, pencil.
Theory: Population refers to total number of individuals of a species found in a given geographical area, sharing
similar resources and can interbreed. Population ecology links ecology to population genetics and evolution. Size
of population depends upon competition with another species, impact of predator or effect of use of pesticides. Size
of population keeps changing. It depends upon Predation, Food availability and weather.
We can study population density by laying quadrats of size 1 sq. m. Quadrat is a sampling plot or area of size 1m x
1m. Chosen randomly as representative of the given area species falling inside the quadrat are recorded to find out
population density.
Procedure:
1. First we will select a quadrat randomly in a uniform area.
2. Using a meter scale, we will make a quadrat of 1m x 1m in the field. We will fix 4 nails at the corners of the
quadrat & tie thread over the nails.
3. We will count the number of plants of each species present in the quadrat.
4. Now we will make other quadrat and note down number of plants in that quadrat also.
Observation:
Name of No. of individual in quadrat Total number of
S.No. Average
plant Spp. I II III IV V individuals of a species
1 A 2 3 0 1 0 6 1.2
2 B 2 1 2 0 1 6 1.2
3 C 4 0 3 2 1 10 2
4 D 3 3 1 2 0 9 1.8

Conclusion:
Population density of a plant spp. is the average number of individual plants of the spp. occurring in an area.
Population Density= Total no. of individuals in quadrat / Total no. of quadrat.
Precaution:
1. In quadrat consider the plant lying under the string if more than half of it lies towards inside.
2. Choose a field having uniform distribution of spp.
3. Count the individuals of one plant spp. at a time.
 EXPERIMENT NO. 3
Aim: Study plant population frequency by quadrat method.
Material Required: Nails, thread, hammer, metre scale, pencil, paper.
Theory: Population frequency may be defined as the number of sampling units in which a given species occurs and
thus expresses the distribution or dispersion of various species in a community.

Population Frequency = No. of quadrats of occurrence of spp. / Total no. of quadrats studied
Divide the quadrat of size 1m x 1m into smaller units because in smaller units individual of different species can be
counted more accurately.
Method:
1. First of all we will select a site and with the help of metre scale make a quadrat of 1m x 1m in the field.
2. Now tie string on the nails.
3. We can divide each quadrat into 16 small squares by tying strings at distance of 25 cm each on either
side. Smaller squares can also be marked with nails and strings.
4. We will note down the number of plants of each spp. in each quadrat.
5. We will find the total number of plants of each spp. in each quadrat.
6. We will select another quadrat randomly and repeat the steps.
Observation:
S.No. Name of No. of individual of spp. Present in Total number of Frequency%
Plant each quadrat quadrats in which spp. =A/5 x 100
spp. I II III IV V Occurred(A)
1 A P P 0 0 P 3 60
2 B P P P 0 P 4 80
3 C 0 P P P P 4 80
4 D P 0 P 0 P 3 60
Conclusion:
1. The frequency percentage for a spp.
= No. of quadrats in which species appeared / Total no. of quadrats studied x 100
2. Frequency of a species gives information about :
a. Types of habitat and correlation of plant with habitat.
b. Interaction of spp. with other species
Precautions:
1. If a plant has more than half of its portion inside the quadrat, it should be assumed and counted inside
the quadrat.
2. Quadrats should be studied from 1 area only, with uniform distribution of plants.
3. The quadrat should be accurately measured.
 EXPERIMENT NO. 4
Aim : Prepare a temporary mount of onion root tip to study mitosis.

Material Required: Onion bulb, conical flask/ glass bottles, corked tube, petridishes, scissors, forceps, needles,
methyl alcohol, acetic acid, hydrochloric acid, acetocarmine, distilled water, spirit lamp, microscope, slides,
coverslips, blotting paper.

Theory: Production of all cells involves the division of pre-existing cells. There are mainly two types of cell
divisions mitosis and meiosis. Somatic cells or body cells are divided by mitosis. The germ cells or reproductive
cells divide by meiosis when they form gamete in animals and spores in plants.
Each cell division consist of two events: karyokinesis and cytokinesis.

Method:
1. First we will take a medium sized bulb of onion and by means of a sharp blade trim off the old roots from
its base.
2. We will place the onion on a conical flask/ glass bottle full of water. Its base should touch the water. We will keep
it for a week to grow the roots.
1. Now we will cut 5mm off the tips of roots and put them into a vial containing a mixture of 1:3 acetic acid
and methanol. Keep for one hour. This process is known as fixation.
2. We will remove 2 or 3 root tips. Hydrolyze them by warming to 600 C in 1 N Hydrochloric acid for 15
minutes.
3. Remove the root tips, wash them thoroughly in water.
4. On a slide place a drop of acetocarmine. We will put one hydrolyzed root tip in a drop and place a coverslip
on the root.
5. We will gently squash the root by tapping the coverslip with the blunt end of a pencil or needle until the
cells separate and spread out into a very thin layer. No air bubbles should be there under the coverslip.
6. We will warm the slide gently over a flame for a few seconds.
7. Now we will observe under microscope to locate the dividing cells.

Observation: Rectangular cells with pink nucleus are seen scattered under low power of the microscope. Under
high power following stages become distinct.
1. Interphase: It refers to non-dividing phase of cell cycle. There is a distinction between nuclear envelope and
nucleolus. Chromatin fibers appear in the form of a network within the nucleus.
2. Prophase: At the end of prophase nuclear membrane and nucleolus start disintegration and disappear.
Chromatin material shortens and condenses into thread like structures known as chromosomes.
3. Metaphase: Each chromosome get attached to the spindle fibers at its centromere. It becomes arranged at
the equator of the spindle. A bipolar spindle develops in the cell and chromosomes become thick and two
chromatids of each chromosome become clear.
4. Anaphase: The two sister chromatids of each chromosome separate from the centromere and move
towards the opposite poles. The daughter chromosomes appear V, J, L and I shapes, depending upon the
position of centromere.
5. Telophase: The spindle disappears, daughter chromosomes uncoil in order to form chromatin fibers at the
two poles. Cytokinesis occurs by cell plate formation between the two daughter nuclei. Nuclear membrane
and nucleolus reappears and two daughter nuclei appear at opposite poles.

Precautions:
1. We should warm slide gently above flame of the spirit lamp.
2. Keep the base of the onion bud in contact of water while growing the roots.
3. Keep root tips in the morning between 8 to 10 a.m.
 EXPERIMENT NO. 5

Aim: Isolate DNA from available plant material.

Material Required: Juice of papaya, 95% ethanol, plant material, mortar and pestle, beakers, test tubes, liquid
detergent, non-iodized sodium chloride, distilled water.

Theory: There are mainly two types of nucleic acids found in living systems i.e. DNA and RNA. By
recombinant DNA technology breeders can introduce foreign DNA in other organisms, including bacteria, yeasts,
animals and plants. Such organisms are known as Genetically Modified Organisms. It involves isolation of DNA
from a variety of sources and formation of new combination of DNA.

Procedure

1. First we will take plant tissue and grind it in mortar by adding 10 ml detergent, salt solution and
filter it through muslin cloth.
2. We will take 10 ml filtrate and add 3-4 ml papaya juice. We will swirl the test tube by holding it
between two hands to mix the contents.
3. Now we will carefully pour 10 ml chilled ethanol down the side of test tube in order to form a layer
on the top of content, let it stand undisturbed for 3 minutes.

Observation
The addition of ethanol to the solution causes DNA to precipitate. The DNA fiber appears as white precipitate of
very fine threads on the glass spool.

Precautions:
1. Thoroughly wash the plant material with distilled water to remove any dust and dried by blotting
before weighing.
2. All the glasswares used must be thoroughly cleaned and dried.
3. Use standard quality of the chemicals and enzymes for the experiment.
 Spotting

Experiment - 1
I. Maize Flower:
1. It is Anemophilus or wind pollinated flower.
2. It is small, unisexual and inconspicuous.
3. They are colourless, odourless and nectarless.
4. Their stigma is hairy feathery or branched to catch pollen grain.

II. Salvia Flower:

1. They are Entomophilus or insect pollinated flower.
2. They are showy or brightly coloured, secrete nectar to
feed insects.
3. They have landing platform for insects.
4. When insects moves in a flower, its head pushes
anther plates & forces fertile anther lobes to strike
against its back.

III. Bigonia Flower:

1. They are ornithophilus or Bird pollinated flower.
2. Flowers are brightly coloured.
3. Floral parts are leathery, secrete abundant nectar.
4. Flowers are odourless or without fragrance.

Experiment - 2
I. Pollen germination on stigma
1. Pollen contains microgametophytes of seed plants.
2. Microgametophytes produce male gametes.
3. They have hard coat that protects sperm cells during
movement.
4. When pollen lands on compatible pistle of flowering plants,
it germinates and produces pollen tube.
 Experiment - 3
I. T.S of testis:
1. It is covered by a thick fibrous tissue called tunica albuginea.
2. They consist of numerous seminiferous tubules
embedded in interstitial tissue.
3. It is lined by germinal epithelium.
4. Sertoli cells are present to provide surface and
nourishment to developing spermatozoa.
5. Interstitial cells or leydig cells are present between tubules,
secrete male sex hormone.

II. T.S of ovary:

1. It is bounded by germinal epithelium.
2. It consist of outer cortex with mature follicle.
3. Its inner medulla consist of rounded or oval graffian follicles.
4. Medulla contains blood vessels, nerve fibres and
smooth muscles.
5. The cortex may also contain corpus luteum.

Experiment No. - 4

I. Meiosis in grasshopper testis through permanent slides:

A. Meiosis I
(a) Leptotene
Chromatin fibres condense and form thick thread like structures called
chromosomes.
Nuclear envelops and nucleolus are distinct.
(b) Zygotene
Homologous chromosomes form pairs called bivalent. This pairing is called
synapsis
The individual of a pair are similar in length and in position of their
centromere.
(c) Pachytene
The two chromatids of each chromosome become visible, so that a bivalent becomes a tetrad in Crossing over
(exchange of chromatid segments between homologous chromosomes) takes place between non-sister
chromatids of homologous chromosomes.
(d) Diplotene
The two chromosomes of each bivalent move away and homologues are held together at one more points called
chiasmata.
(e) Diakinesis
Homologous chromosomes appear thick and ring shaped.

2. Metaphase I
Nucleolus and nuclear envelope disappear and spindle begins to be formed. The bivalent (homologous
chromosomes) arrange themselves at the equator of the spindle.
The spindle get attached to the centromere of the chromosome.
3. Anaphase I
The two chromosomes of each bivalent move to the opposite pole. The two Each pole has half the number of
chromosomes with two chromatids each.
4. Telophase I
The chromosome at each pole uncoil, and nucleolus and nuclear envelope reappear.
Cytokinesis occurs to form two haploid daughter cells.
Interkinesis
A very short interphase may intervene between meiosis I and meiosis II.

B. Meiosis II
It includes following four stages:
1. Prophase II
The chromosomes of daughter cell begin to condense and become thick.
Nuclear envelope and nucleolus begin to disappear.
2. Metaphase II
The chromosomes are arranged on the equator of the spindle.
Each chromosome is held by the spindle at the centromere to both the poles.
3. Anaphase II
The sister chromatids (daughter chromosomes) of each chromosomes separate and migrate towards the opposite
poles.
Each pole thus, receives haploid number of chromosomes.
4. Telophase II
The chromosomes begin to uncoil and become thin.
The nuclear envelope and nucleolus are reconstituted.
Cytokinesis occurs and four daughter cells are formed, each with haploid number of chromosomes.

Experiment No. - 5

I. T.S of blastula:
1. It is a 32 cell stage of embryo development.
2. Its outer cell layer becomes flattened and form a layer
called trophoectoderm.
3. Its remaining cell form mass of cell called inner cell mass
leaving a cavity aside called blastocoel.
4. The inner cell mass is a precursor of embryo.

Experiment No. - 6
I. Study of Mendelian inheritance using seeds of different colours/ size of any plant:
1. The contrasting forms in both the traits of pea seed (e. seed shape and seed colour) show an
approximate ratio of 3: 1.
2. This ratio is exactly the same as obtained by Mendel for monohybrid crosses and indicate that the
dominant and recessive forms of seed shape and seed colour exist in the ratio of 3: 1 in the population of
pea seeds.
Total no. of No. of seeds showing
S.No. Characters Approx. ratio
seeds observed contrasting trait
Seed Shape
1. 106 80 (Round) : 26 (wrinkled) 3.07 : 1
(Round/Wrinkled)
2. Seed Colour (Yellow/Green) 110 83 (Yellow) : 27 (Green) 3.07 : 1

Experiment No. - 7
I. Rolling Tongue – Pedigree Chart
Problem: Inability to roll the tongue appears in the progeny due to recessive
gene. Find out the possible genotype of the family members in the
following pedigree.

Solution:
1. The trait is present in father due to presence of two recessive genes (I-2 aa).
2. Trait can appear in progeny only when it becomes homozygous recessive.
Since only one of the progeny carries the trait, the mother parent must be
heterozygous(I-1Aa)
3. The cross between II-1 and her husband also produces one homozygous
recessive (III-2aa). This is possible only if the outsider is heterozygous
(Aa).
4. II-3 is heterozygous (Aa). The new entrant in the pedigree is homozygous
 dominant (AA).

II. Widow’s Peak- Pedigree Chart

Problem: In the pedigree given below, indicate whether the shaded
symbols belong to dominant or recessive trait. Also give genotype
of the whole pedigree.
Solution:
1. Since the shaded symbol appears in all offspring, father must be
homozygous dominant(AA) while mother homozygous recessive.
2. All the members of second generation will be heterozygous (Aa).
This is confirmed by marriage of II-1 with homozygous recessive
bears children of both parental types
3. Marriage of II-3 with the homozygous recessive can produce both
recessive and heterozygous .

Experiment No. – 8

I. Emasculation
Comments
(i) This method is employed in the crops having
flowers of sufficiently large size lint cotton.
(ii) The instruments used in this method include
pocket lens, forceps, needle, scissors, scalpel, camel
hair brush etc.
(iii) In this process anthers are removed from the
flowers before their maturation.
(iv) The anthers are cut with the help of sterilized
forceps or scissors.

II. Bagging and tagging.

Kommentare
(i) After emasculation, the flowers are covered with small
bags to prevent pollination with undesired pollen grains.
(ii) These bags are made of polythene, paper, muslin cloth
or parchment paper.
(iii) The bags are punctured or made perforated so as to
provide aeration to the flowers.
(iv) The flowers of male parents are also protected in bags
to prevent mixing of their pollen grain with foreign
pollens.
(v) After dusting of the desired pollen grains on the
emasculated flowers, the bags are retagged.
(vi) A label of paper is tagged on the plant which displays
the date of emasculation, crossing and brief account of the
parents.
 Experiment No. - 9
I. Ascaris:
1. It is Ascaris lumbricoides.
2. It causes Ascariasis.
3. Patient feel abdominal discomfort and Colic pains.
4. Patient suffers from diarrhoea, vomiting and slight temperature increase.
5. They may block the intestine and appendix.

II. Entamoeba:
1. It is Entamoeba histolytica.
2. It causes Amoebic dysentery, Amoebiasis.
3. As a result of infection patient passes out stool with blood
and mucus and suffers from abdominal pain, nausea,
flatulence and bowel irregularity with headache.
4. Entamoeba feeds on RBC by damaging wall of large
intestine and reaching the blood capillaries.

III. Plasmodium:
1. It is Plasmodium vivax.
2. It causes malaria.
3. Symptoms appear after 14 days of infection.
4. Patient suffers from chills, shivering and high temperature.
5. The patient become anaemic.

IV. Ringworm:
1. It is caused by Trichophyton species.
2. It causes disease Athlete’s foot.
3. It forms lesions on skin.
4. It also infects nails of hand and feets.
5. As a result skin becomes dry & whitish in colour.
 Experiment No. - 10
I. Lichen
Comment:
1. Lichens are composite organisms representing a symbolic association between a fungus & an alga.
2. They grow on lands, rocks, tree trunk & wall of houses, like dry vegetation.
3. The thallus of lichen resembles neither alga nor fungus.
4. Lichen reproduces vegetatively by fragmentation, asexually by soredia & isidia.
5. Sexual organs like those in Ascomycetes are formed.

Experiment No. - 11
1. Homologous Organs in Plants
(i) Tendrils of passion fruit and thorns of pomegranate
Tendrils of passion fruit and thorns of pomegranate are structurally and functionally different but they have similar
origin i.e. they arise from axillary bud.

2. Analogous Organs in Plants

(i) Stem tendrils and leaf tendrils. All tendrils are analogous with one another, being structurally and functionally
similar, irrespective of their origin. Example: Tendrils of pea and tendrils of Vitis. Tendrils of pea are modification
of leaf and in Vitis it is the modification of terminal bud.

3. Homologous Organs in Animals

(i) Wings of birds, and forelimb of mammals/reptiles/frog: All have the same bony elements (humerus, radio-ulna,
carpals, metacarpals and phalanges), but perform different (flying in birds, for holding or walking etc. in other)
functions.

4. Analogous Organs in Animals

(i) Wings of dragonfly/cockroach/butterfly and of birds: Wings of insects and wings of birds are different in origin
and structure but perform similar function.