Thesis report on Genetic study of Bhumij Tribe of Jharkhand using mt DNA and Y chromosome DNA marker by Smita Bernadet Kujur - HTML preview

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Chapter IV: Analysis of Results

4.1 Discussion of Results

4.2.1 Y-chromosomal Analysis

4.2.2 Mitochondrial DNA Analysis

Chapter V : Summary and Conclusion

4.3 Summary and Conclusion

Terms

Bibliography

LIST OF FIGURES

Fig 1 : Human Mitochondrial DNA

Fig 2 : Map of human haplotype migration, according to mitochondrial DNA

Fig 3 : mt DNA haplogroup distribution of world

Fig 4 : Structure of Y chromosome

Fig 5: Major Haplogroup Frequencies Of the World

Fig 6: The different transmission paths of genetic material. Y-chromosomes

exclusively paternal, mitochondrial DNA entirely maternal.

Fig 7: Out-of-Africa model

Fig 8: People who contributed

Fig 9: Vacutainer

Fig 10 : Transfering blood from syringe to vacutainer

Fig11 : MJ Research PCR

Fig 12 : PCR ( Eppendorf and Veriti)

Fig 13: DNA Sequencer

Fig 14: DNA sequencing analysis software

Fig 15: Auto Assembler Software

Fig 16 : aqueous layer, protein layer and solvent layer.

Fig 17 : 2 clear layers of DNA and Chloroform .

Fig 18 : DNA Extracted

Fig 19 : Gel Check of Dilution

Fig 20 : Gel Check of PCR products

Fig 21 Map: Site of sample collection

Fig 22 : Consent Form

Table 1

Fig 23 : Frequency Chart of Y haplogroup

Table 2

Fig 24 : Frequency Distribution of mt DNA

Figure 25. Worldwide frequency distribution of Haplogroup O.

Figure 26. Relative frequency distribution of the four main subclades of

Haplogroup O.

Fig 27: Derived samples derived from M95 primer leads to O2a‐

Haplogroup On Y chromosome phylogenetic tree

Fig 28: Derived samples derived from M82 primer leads to H1‐

Haplogroup On Y chromosome phylogenetic tree

Fig 29 : A‐G Mutaion

Fig 30: Insertion T

Fig 31 : M82 primer haplogroup analysis giving Derived

Fig 32 : M95 primer haplogroup analysis giving Ancestral

Fig 33 : M95 primer haplogroup analysis Giving Derived

LIST OF ABBREVIATIONS

%

-

Percentage

C

-

Degree Celsius

ATP

-

Adenosine 5’-triphosphate

bp

-

base pair(s)

cm

-

centimeter

cpm

-

counts per minute

dATP

-

2’-deoxyadenosine 5’-triphosphate

dCTP

-

2’-deoxycytidine 5’-triphosphate

DDW

-

Double distilled water

dGTP

-

2’-deoxyguanosine 5’-triphosphate

D-loop - The displacement loop

DNA

-

Deoxyribonucleic acid

dNTP

- 2’-deoxynucleotide 5’-triphosphate

ddNTP

- 2’,3’-dideoxynucleotide 5’-triphosphate

dTTP

-

2’-deoxythymidine 5’-triphosphate

EDTA -

Ethylene diamine tetra acetic acid

Et.Br

-

Ethidium bromide

Extn

-

Extension

Figure

-

Figure

g

-

gram

kb

-

kilo base

M

-

molarity

mA

-

milli ampere

mg

-

milligram

min

-

minutes

ml

-

millilitre

mm

-

millimeter

mM

-

millimolar

mtDNA

-

mitochondrial DNA

mtRNA - mitochontrial RNA

rRNA - ribosomal RNA

tRNA - transfer RNA

N

- Normality

nm

- nanometer

NaOH

-

sodium hydroxide

ng

- nanogram

OD

-

Optical density

OH - Origin of heavy chain replication OL - The L-strand origin PCR

-

Polymerase chain reaction

pM

- picomole

RNA - ribonucleic acid

rpm.

-

Revolutions

per

minute

SDDW

-

Sterile Double Distilled water

SDS

-

Sodium dodecyl sulphate

Sec.

-

Seconds

SNPs - Single Nucleotide Polymorphisms

SRY

-

Sex-Determining Region On Y Chromosome

SSC

-

Sodium

saline

citrate

STR

-

Short

Tandem

Repeat

TAE

-

Tris-Acetate-EDTA

TE

- Tris-EDTA

Tris

-

Tris

( ecogniz methyl) amino methane

TMRCA - Time to the most recent common ancestor

U

-

unit

UEP - unique event polymorphism

UV

- Ultraviolet

V

- Volts

v/v

- Volume/Volume

w/v

- Weight/Volume

g

- Microgram

l

- Microlitre

MW -

Micro molar Watts

YAP

-

Y- Alu polymorphism

YCC - Y Chromosome Consortium

ABSTRACT

India is a conglomeration of various ethnicities with 4693 communities, 325

languages, 25 scripts and numerous endogamous groups. It is a home of several

tribal pockets, which represents different genetic isolates and thus provides unique wealth to understand human evolution. These autochthonous tribal populations

reveal striking diversities in terms of language, marriage practices as well as in

their genetic architecture. The origin and settlement of the Indian people still

remain intrigues for the scientist studying the impact of the past and modern

migration of the genetic diversity and structure of contemporary populations.

Indian populations are stratified as tribe, caste and religious community.

Endogamy has probably been a major reason for genetic diversification among the

people of this region. Taking geographical and ethnic diversity into account and to answer the question of origin and evolution of maternal and paternal lineages

of Indian population .Above 400 base pairs of the HVR-1 region and selected

coding regions of the mitochondrial DNA (mtDNA) and Y chromosome markers

in 102 individuals of Bhumij, an Austro-Asiatic tribe of Jharkhand, was

analyzed and compared with the data available from the Indian subcontinent.

Based on the mutations observed in the HVR -1 and selected coding region of

mitochondrial DNA, haplogroups were assigned to each of the individual. It was

observed that most of the individuals of Bhumij tribal population were falling in

Indian specific macro haplogroup M, displaying the array of South Asian specific

lineages. On the other hand, Y chromosomal analysis is showing 70% percentage

of individuals falling into O2a-M95 haplogroup, found frequently among Austro-

Asiatic. Moreover, it is evident that our investigation of the small population is a snapshot with respect to the peopling of the Indian subcontinent. In future,

detailed phylogeographic and phylogenetic analysis of more tribal population can

reveal the detailed account of maternal and paternal lineages as well as genetic

affinity of the Indian population.

Chapter 1