P.S.Chidambara Nadar Senior English School, Virudhunagar.

Lab Manual class XII Biology

Exp No 1: Isolation of DNA
Aim: To isolate DNA from available plant material such as Onion, fresh green pea seeds, green papaya, etc.
Requirements: Plant materials, mortar and pestle, beakers, test tubes, ethanol, etc.
Procedure:
 Take a small amount of onion and grind it in a mortar with a little amount of water and sodium chloride.
 Make it into a solution and filter it.
 To this filterate, add liquid soap solution or any detergent solution and mix it with a glass rod.
 Then tilt the test tube and add chilled ethanol and leave it aside in the stand. After few minutes we can observe the precipitated
DNA as fine threads.
 DNA that separates can be removed by spooling
Observation: DNA appears as white precipitate of very fine threads on the spool. Inference: Thus DNA can be isolated from the
plant cell nucleus by this technique.
Precautions:
1. All the glasswares must be thoroughly cleaned and dried.
2. The chemicals used for the experiments must be of standard quality.
3. If ordinary ethanol is used, the time duration for obtaining precipitated DNA may extend further.
Exp No 2: Study of pollen germination
Aim : To study of pollen germination on a slide.
Requirements: Fresh seasonal flowers, slide, coverslip, microscope, sucrose, boric acid, magnesium sulphate, potassium nitrate,
beakers etc.
Procedure:
1. Prepare a nutrient solution by dissolving 10g of sucrose, 10mg of boric acid 30mg of magnesium sulphate, and 20 mg of
potassium nitrate in 100 ml of water.
2. Take a few drops of this solution on a slide, and dust a few pollen grains from the stamen of a mature flower on it.
3. Observe the slide in the microscope after 5 minutes and then observe it regularly for about half an hour.
Observation : In nutrient medium pollen grains germinate. The tube enlarges and comes out of the pollengrain through one of the germ
pores to form a pollen tube. Pollen grains different plant species show different types of sculpturing. Rate of pollen germination and
viability also various for different species.
Result : Pollen grains from different plant species take different time for their germination.
Precautions:
1.Flowers should be freshly plucked .
2. Use clean slide to observe the pollen grains
Exp No 3: Study of Pollen germination on a slide
Aim: To study pollen germination on a slide.
Materials Required: Permanent slide of pollen germination, microscope.
Procedure: Fix the permanent slide under the microscope, Observe the slide under the low power and then under high power of the
microscope.
Observation:
 A number of germinated pollen grains are seen . A thin tube emerges out of each pollen grain.
 Each pollen grain has a prominent two layered wall.
 The hard outer layer called the exine is made up of sporopollenin and has prominent aperture called germpore.
 The inner wall of pollen grain is called intine which is made up of cellulose and pectin.
 Intine forms the pollen tube that comes out through one of the germ pore.
 Pollen tube shows two small male gametes and a tube nucleus which is disintegrating.
Precautions:
1. Do not disturb the slide
2. First focus the slide under low power and under the high power microscope
Anaemophilous flower Exp No 4: Floral adaptation to pollination by different agents
Aim: To study the flowers adapted to pollination by different agencies (wind, insects and birds)
Requirements:Fresh flowers of maize or any other cereal / grass, salvia/ Ocimum and Brassica (mustard), forceps, hand lens, slide,
needle etc.
Procedure: Place the given flower on the slide and observe it with the help of a hand lens. Note down the adaptations of the
flowers meant for pollination by the external agencies.
Maize flowers (Anemophillous or wind pollinated flowers)
The flowers of maize show following adaptations for pollination by wind
1. The maize plant is monoaciou sand bears unisexual flowers. The male flowers are borne in terminal inflorescence while
the female flowers are borne in axillary inflorescence.
2. Flowers are small and inconspicuous.
3. The flowers are colour less, odourless and nectarless.
4. Flowers are produced above the foliage or placed in hanging position.
5. Both stigmas and anthers are exerted.
6. Anthers are versatile, and the pollen grains are light, small and dusty.
7. The pollen grains are produced in very large numbers.
Entamophilous flower Salvia flowers: (Entamophilous or insect pollinated flowers)
The flowers of salvia show following adaptations for pollination by insects.
1. The flowers are showy or brightly coloured for attracting pollinating insects.
2. The flowers are borne in verticellaster inflorescence to become conscipcuous.
3. Flowers secrete nectar to feed visiting insects. Nectar glands are placed in such a position that an insect must touch both
the anther and stigmas
4. The flowers landing platform for the insects
5. The flowers are protoandrous with bilipped corolla and have turn pipe or lever mechanism.
6. Each stamens has long connective which bears a fertile anther lobe at the upper end and the sterile plate like another
lobe at the lower end.
Ornithophilous flower
Bignonia / Callistemon (bottle brush) flowers (Ornithophilous or bird pollinating flowers)
The flowers of Bignonia shows the following adaptations :
1. The flowers are usually brightly coloured –red, orange yellow or blue.
2. The floral parts are commonly leathery.
3. In some cases, the corolla are leathery.
4. Flowers secrete abundant watery nectar or have edible parts.
5. The nectar is secreted in such abundance that drops of it can be brought down by shaking branches.
Emasculation 6. The flowers are generally odourless or without fragrance.

Exp No 5: Controlled pollination – Emasculation, bagging and tagging

Aim: To comment on the exercises of hybridisation (emasculation, bagging and tagging) through models/ charts.
1.Emasculation:
Identification: Forceps or scissors method of emasculation
Comment: 1.This method is employed in the crops having flowers of sufficiently large size (eg. lint cotton).
2. The instrument used in this method include pocket lens, forceps, needle, sciccors, scalpel, camel hair brush etc
3.In this process, anthers are removed from the flowers before maturation.
4.The anthers are cut with the help of sterilized forceps or scissors
Bagging and Tagging: 2.Bagging and Tagging:
Identification: Bagging and tagging and labelling.
Comment: 1.After emasculation the flowers are covered with small bags to prevent pollination with undesired pollen grains.
2.These bags are made up of polythene, paper, muslin cloth or parchment paper.
3.These bags are punctured or made perforated so as to provide aeration to the flowers.
4.The flowers of male parents are also protected in bags to prevent mixing of their pollen grain with foreign pollens.
5.After dusting of the pollen grains on the emasculated flowers the bags are retagged.
6.A label of paper is tagged on the plant which displays the date of emasculation, crossing and brief account of the parents.
Exp No 6: Study of T.S of testis and ovary
T.S of Testis Aim
To identify the stages of gamete development, i.e. T.S. of testes and ovary through permanent slides from mice.
Materials Required
Permanent slides of T.S. of testes.
Permanent slides of T.S. of the ovary.
Observation
T.S. of Testes
The testes comprise several seminiferous tubules embedded in the interstitial tissues.
Thick fibrous tissues called tunica albuginea cover the testes.
It comprises different types of cells from the outside towards the lumen in the following sequence given below given below:
Spermatogonia → Spermatocytes → Spermatids → Spermatozoa (sperms)
Sertoli cells are located between the germinal cells.
T.S. of Ovary A large number of spermatozoa with their heads embedded in the sertoli cells are present in the lumen of the seminiferous tubule.
The Leydig cells that produce testosterone are present in the interstitial tissues.
T.S. of Ovary
An ovary is a germinal epithelium bounded by a solid structure covered by a thick layer of fibrous tissue known as tunica albuginea.
It consists of an inner medulla and an outer cortex.
The medulla comprises several round or oval bodies known as ovarian follicles.
Follicle development takes place in the following stages:
1°follicle → 2°follicle → 3°follicle → Graffian follicle → Corpus luteum
Cortex comprises corpus luteum along with mature follicles.
The medulla consist of the blood vessels, nerve fibres and some smooth muscles.
T.S. of Blastula Exp No 7: Study of Blastula
Aim : To observe the TS of mammalian blastula through permanent slides.
Materials Required
Permanent slide, Compound microscope
Procedure
Observe the permanent slide under a microscope and note down the features of T.S of the mammalian blastula.
Observations
 The blastula is a spherical mass of about sixty four cells.
 It has a fluid filled cavity known as blastocoel.
 An outer layer of blastomeres known as trophoblasts is observed.
 One end of the blastula shows a cellular mass adhered to the trophoblast. This is known as the inner cell mass.The inner cell mass is the
precursor of the embryo
 Exp No 8 : Temporary mounting of the onion root tip.
Aim : To prepare temporary acetocarmine stained mount of onion root tip to study various stages of mitosis
Requirements: Onion bulbs, conical flasks/glass bottles, petridishes, scissors, foreceps, needles,and alcohols acetic acid, acetocarmine, distilled water,
Bunsen burner, microscope, slides, coverslips, blotting paper.
Procedure:
Take an onion and place it on the tile.
Carefully remove the dry roots present using a sharp blade.
Grow root tips by placing the bulbs in a beaker filled with water.
New roots may take 3–6 days to grow.
Cut off 2–3 cm of freshly grown roots and let them drop into a watch glass.
Using a forceps, transfer them to the vial containing freshly prepared fixative of aceto-alcohol (1:3: glacial acetic acid: ethanol).
Keep the root tips in the fixative for 24 hours.
Remove 2 or 3 root tips and hydrolyse them by warming to 60o in 1 N hydrochloric acid for 15 minutes.
Place a drop of acetocarmine on a slide. Put one hydrolysed root tip in the drop and place a coverslip over it.
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.
Warm it slightly on burner. Care should be taken that the stain is not dried up.
Carefully blot the excess stain using filter paper.
This preparation of onion root tip cells is now ready for the study of mitosis.
Place the slide under the compound microscope and observe the different stages of mitosis.
Various stages of mitosis are prophase, metaphase, anaphase and telophase.
Events during Mitosis
Prophase: 1. Mitosis begins at prophase with the thickening and coiling of the chromosomes.
2. The nuclear membrane and nucleolus shrinks and disappears.
3. The end of prophase is marked by the beginning of the organization of a group of fibres to form a spindle.
Metaphase : 1. The chromosome become thick and two chromatids of each chromosome become clear.
2. Each chromosome attaches to spindle fibres at its centromere.
3. The chromosomes are arranged at the midline of the cell.
Anaphase : 1.In anaphase each chromatid pair separates from the centromere and move towards
the opposite ends of the cell by the spindle fibres.
2. The cell membrane begins to pinch at the centre.
Telophase : 1. Chromatids arrive at opposite poles of cell.
2. The spindle disappears and the daughter chromosome uncoils to form chromatin fibres.
3. The nuclear membranes and nucleolus re-form and two daughter nuclei appear at opposite poles.
4. Cytokinesis or the partitioning of the cell may also begin during this stage.
Exp No 9: Study of Meiosis using Onion floral bud
Aim : To observe the stages of meiosis on onion bud cells through permanent slides.
Materials Required
Permanent slides of meiosis
Compound Microscope
Procedure
1. Place the slide on the stage of the microscope.
2. Observe it under low power and then under high power..
Observations
The different stages of meiosis are observed along on the basis of the following features.

Stages of Meiosis I
Prophase I
In this stage, the chromosomes condense and move towards the centre of the cell. It consists of five different sub-phases:
Leptotene: the chromatin fibres condense and form thick thread like structures called chromosomes. Nuclear envelope and nucleolus are distinct
Zygotene: Synapsis between homologous chromosomes start. The chromosomes pair to become bivalents
Pachytene: the bivalents become tetrads. Crossing over takes place between the non-sister chromatids of the homologous chromosomes.
Diplotene: The two homologous chromosomes migrate apart and homologous are held at more than one part called chiasmata.
 Diakinesis: The nucleolus and the nuclear envelop disappear at this stage and the centrosome moves to the equator.
Metaphase I
The homologous chromosomes that contain two different alleles for each gene, line up on the metaphase plate to be separated. The spindle gets attached
to the centromere of the chromosome
Anaphase I
The two chromosomes of each bivalent move to the opposite pole. Each pole has half the number of chromosomes with two chromatids each.
Telophase I
The chromosomes are completely pulled apart and new nuclear envelope forms.
Stages of Meiosis II
Prophase II
In this stage, the chromosomes become thick and condensed , the nuclear envelope disintegrates and centrioles develop.
Metaphase II
The chromosomes line up on the metaphase plate and the chromatids are on either side of the metaphase plate.
Anaphase II
The sister chromatids separate and are known as sister chromosomes. Each pole receives half the number of the chromosomes.
Telophase II
The chromosomes begin to uncoil and become thin. The nuclear envelope and nucleolus are reconstituted.
Cytokinensis: The cells divide and four daughter cells are formed, each with haploid number of chromosomes
Exp No 10: Mendelian inheritance using seeds of different colour/ sizes of any plant.
Aim: To study Mendelian inheritance using seeds of different colour/ sizes of any plant.
Requirements: Pea seed sample, enamel tray, petridishes, notebook pencil.
Procedure: 1.Take a lot of about 100 pea seeds in an enamel tray.
2.Separate out round and wrinkled seeds and put them in separate petridishes.
3.Notedown the number of round and wrinkled seeds and calculate their approximate ratio.
4.Repeat the process for the other contrasting traits of the seed ie, yellow and green colour.
Conclusion: The contrasting forms in both the traits of pea seed ( colour and size of seed), show an approximate ratio of 3:1. This ratio is exactly the
same as obtained by Mendel for monohybrid crosses and indicate the dominant and recessive forms of seed shape and seed colour exist in the ratio of
3:1 in the population of pea seeds.
Exp No 11 : Pedigree chart analysis
Aim: To study the prepared pedigree charts of genetic traits such as rolling of tongue,blood groups, widows peak, colour blindedness etc.
Requirements: Prepared pedigree charts of the genetic traits
Procedure: Observe the pedigree chart and write the comment on it.
1. Autosome Linked Dominant traits: These are the traits whose encoding gene is present on any one of the autosomes, and the wildtype allele is
recessive to its mutant allele, i.e., the mutant allele is dominant.
The characteristic features of inheritance of such type of traits are:
he characteristic features of inheritance of such type of traits are:
(a) Transmission of traits occurs from parents of either sex.
(b) Males and females are equally affected.
(c) The pedigree is vertical, i.e., the trait is marked to be present in each of the generations.
(d) Multiple generations are characteristically affected.
(e) Never transmitted from parents who donot posses the trait.
Brachydactyly, polydactyly, dimple in the cheek and widow's peak are some of the common traits of this type.
Widow’s Peak
It is a hair line that forms distinct peak as it crosses the forehead. It is a dominant trait .Thus homozugous dominant and heterozygopus individuals have
widow’s peak whereas homozygous recessive individuals have straight hair line.
Autosomal Recessive trait: These are the traits whose mutant allele is recessive to its wild type allele.
The following are the salient features of the inheritance of such type of traits.
(a) Occur in equal proportions in multiple male and female siblings, whose parents are normal but carriers;
(b) The siblings are homozygous for the defective allele, but their parents, though some may appear normal, are obviously heterozygous, i.e., are merely
carriers of the trait.
(c) Consanguinity (marriage between man and woman genetically related to each other, such as cousins) occasionally results in the appearance of such
traits.
Rolling of Tongue:
 It is the ability of a person to roll the tongue in U shape. The ability to roll the tongue is caused by autosomal recessive allele ‘a’. Thus both homozygous
dominant(AA) and heterozygous (Aa) individuals are able to role the tongue, while homozygous recessive individuals are unable to roll the tongue.
X-linked Recessive traits: These are the traits whose encoding gene is present on the X-chromosome and its mutant allele is recessive to its wild-type
X-linked Recessive traits:
allele. Red-green colour blindness and hemophilia, are some of its well known examples.
The characteristic features of such inheritance are:
(a) Females express the trait only when they are homozygous for the mutant allele, whereas the males do so even when they are hemizygous for it
(b) The males could never provide the trait their son as the gene is present in the X chromosome.
(c) The grandsons get the trait from their grandmother hence skipping of generations takes place.
Colour Blindness: Colour blindness is a sex linked disorder in which the affected individual is not able to differentiate between red and green colours.
Affected individuals are relatede to one another through the maternal side of their family. The traits are often noticed in the males as the Y chromosme
does not have the gene counterpart that is present in the X chromosome. Females may be healthy as they have two X chromosomes. The heterozygous
females are carriers.
Exp No 12 : Study of disease causing organisms
Aim: To study the disease causing organisms through permanent slides and specimens.
Ascaris
Phylum: Aschelminthes
Class: Nematoda
Type: Ascarislumbricoides
Ascarisl umbricoides Ascaris exhibits the following characteristic features:
1. It has a long, cylindrical and unsegmented body.
2. The male and female organisms are separate.
3. It bears a mouth at the anterior end surrounded by three lips.
4. There is an excretory pore on the ventral surface slightly behind the anterior end.
5. A pair of penial spicules are present in the male worms close to the cloacal opening.
6. The female genitals are present at about one-third distance from the anterior end.
Ascariasis is the disease caused by Ascarislumbricoides or roundworm.
Symptoms:
Abdominal cramping
Abdominal swelling
Nausea
Vomiting
Fever
Entamoeba
Phylum: Protozoa
Class: Rhizopoda
Type: Entamoeba hystolytica
Following are the characteristic features of Entamoeba:
1. It is a unicellular organism with an irregular shape.
2. It consists of a few food vacuoles. The contractile vacuole is absent.
3. Cysts with four nuclei are present.
4. It consists of a nucleus located eccentrically in the cell.
Entamoeba histolytica is an organism found in the intestines of humans that is responsible for causing amoebic dysentery.
Symptoms:
Abdominal pain
Watery diarrhoea with mucus, blood and pus
Fatigue
Fever
Nausea
Vomiting
Plasmodium
Phylum: Protozoa
Class: Sporozoa
Type: Plasmodium vivax
Plasmodium can be identified by the following characteristic features:
 1. It is a unicellular endoparasite found within the red blood cells of the diseased person.
2. The parasite is mostly diagnosed at the “signet ring” stage where the parasite appears as a round body.
3. There is a big vacuole present inside the cell. The cytoplasm is accumulated at one place and contains the nucleus.
Plasmodium vivax is a protozoan parasite that causes malaria in humans. The infected female anopheles bites a healthy person and transmits the
sporozoite into the peripheral blood vessels of humans, thereby, causing malaria.
Symptoms:
High fever , Shaking chills from moderate to severe., Headache, Vomiting, Nausea
Ringworm
Kingdom: Fungi
Class: Deuteromycetes
Type: Trichophytonrubrum
Trichophyton or ringworm fungus has the following characteristic features:
1. This fungus feeds on the keratin of the skin of human beings.
2. The hyphae are waxy and can be smooth or cotton-like.
3. Hyphae that are not stained are yellowish-brown, reddish-brown or white in colour.
Ringworm is a communicable fungal infection of the skin.
Symptoms:
Scaly, itchy skin
Red and raised patches
They are redder at the periphery than at the centre and forms a ring-like appearance.
Exp No 13: Study of population density of plants by quadrant method.
Aim :To study the plant population density by the quadrant method.
Materials Required:Field, Nail, thread, metre scale, hammer.
Procedure:
1. Select a site for the study and hammer the nails on the site without harming the vegetation.
2. Fix four nails in the form of a square.
3. Each end of the nail is tied with the help of a thread making a 1m*1m quadrant.
4. Nine more similar quadrants are made at the site of the study.
5. The number of individuals of species A present in the first quadrant is counted and the data is recorded in the table.
6. The number of individuals of species A in other quadrants is also counted and the data is recorded in the table.
7. Similarly, count the number of individuals of species B and C present in all the quadrants and record the data in the table.
8. The density of the plant population is then calculated by the following equation:
Density, D = Total number of individuals of the species in all the sampling units (S)
-------------------------------------------------------------------------------------
Total no of sampling units studied (Q)
Result: The population density of few plant species are more in number and few plant species are less in number and very few are in average number in
Observations
the given area.
Precautions: 1. Measure the quadrat accurately.
2. Mark the quadrats close to each other from one field only.
3. The vegetation should not be damaged while laying the quadrat.
Exp No 14 : Study of the plant population frequency by the quadrant method.
Aim :To study the plant population frequency by the quadrant method.
Materials Required: Field, Nail, thread, metre scale, hammer.
Procedure
1. Select the site of study and make a quadrant of 1m*1m using the nails and the thread.
2. Fix the nails with the help of a hammer without destroying the vegetation.
3. Make nine similar quadrants at the site of study.
4. The plant species for the study should be selected.
5. Observe the species in the first quadrant and mark them as species A.
6. Check the presence of species A in all the quadrants and record the observations in the table.
7. Similarly, record the number of species B and C in all the quadrants and mention them in the table.
8. Determine the frequency of plant population by the formula:
Percentage frequency= (No. of sampling units in which species occur)/(Total number of sampling units used in the study)*100
Result: The plant species ……. Shows the maximum percentage frequency representing the most frequent species. The ………. Shows minimum
percentage frequency representing the least frequent species. (rare species of the given area)
Precations: 1.Measure the quadrat accurately.
 2.Record the presence and absence of different species carefully.
homologous organs in animals Exp No 15 : Study of Homologous organs in animals and plants

Aim: To identify and study homologous organ in animals

Materials required: Charts showing bone structure of hand of man, flipper of whale/ seal, wings of birds, wing of bat and forelimb of cheetah
Observation:
Wings of birds, and forelimb of mammals/reptiles/ frog: All have the same bony elements (humerus radius, ulna, carpals, metacarpals and phalanges),
but perform different (flying in birds, for holding or walking etc. in other) functions.
Conclusion: since all the above animal organs have similar set of bones ,we conclude that they have common origin. Hence they represent homologous
organs though they have different functions.
Homologous organs in plants
Homologous organs in plants Aim: To identify and study the homologous organs in plants
Materials required: Specimens or charts of pea and barberry leaves
Observation:
Tendril of a pea plant and spines of barberry
1.Tendril of pea plant is a modified leaf and is used for providing mechanical support
2. Spines of barberry are also modified leaves, but are meant for protection.
Conclusion:
Since tendril of the pea plant and spines of barberry have a common origin (both are leaves). They represent homologous organs in plants, though they
have similar functions.
Exp No 16 : Study of analogous organs in animals and plants
Aim: To identify and study the analogous organs of animalsandplants.
Materials required:Charts showing the structure of wing of insects, birds and bat.

Analogous organs in animals Observation:

The wing is the organ present in three different organisms that perform the same flying function but are different in structure. In birds, the wing structure
is formed of bones covered with feathers. The wings of insects are nothing but the extension of the integuments, which is the outermost layer of any
animal that protects the body. In bats, the wings are simply the folded skin of their fingers.
Since the wing of insect, birds and bats have common
Analogous organs in plants
Aim: To identify and study the analogous organs in plants.
Analogous organs in plants Materials required: The stem of opuntia and leaves of peepal and potato and sweet potato
Observation:1.The stem of opuntia and leaves of peepal are analogous organs in plants. In opuntia the stem is modified into a broad succulent leaf like
structure that performs photosynthesis like leaves. Peepal leaf is a normal leaf that performs photosynthesis.
Conclusion
Thestem of opuntiaandleavesofpeepal have different origin but perform the same function of photosynthesis they are analogous organs.
2. Potato and sweet potato are examples of analogous organs. Both sweet potato and potato have a common function i.e., storage of food in the form of
starch. Hence, they show convergent evolution. But they both differ in structural origin as the sweet potato is a root modification while the potato is a
stem modification.
Potato and sweet potato have different origin but have common function of storage of food they are analogous organs.

Exp No 17: study of symbiotic and parasitic association in different organisms

Aim: To study the symbiotic and parasitic association in different organisms
Materials required: Specimen of Root nodules of Leguminous plants and specimen of lichens
Root of leguminous plant Lichens Observation: Symbiotic relationship between leguminous plant and Rhizobium bacteria
1. Leguminous plants belong to the family fabaceae
2. These plants have a symbiotic relationship with the soil bacteria called Rhizobium.
3. These bacteria live in the roots of these plants forming root nidules and fix atmospheric nitrogen into nitrates and nitrites for
the plants
4. Plants provide nutrition to the bacteria.
Lichens
1. In the lichens the interaction between algae and fungi is a symbiotic relationship.
2. Algae provide food to fungi that lack chlorophyll pigments.
Cuscuta and host plant 3. Fungi on the other hand help algae by providing protection and aiding in water absorption.
Parasitism (Cuscuta and host plant)
Cuscuta is a parasitic plant which develops special roots called haustoria.
It winds around plants like tomato,tobacco,clover etc. and penetrates the host stem via haustoria
Thus it forms direct connections to vascular bundles of their host to withdraw water, carbohydrates and other solutions.