CONTENTS
Page
ACKNOWLEDGEMENTS iii
ABSTRACT iv
LIST OF TABLES ix
LIST OF FIGURES xi
LIST OF ABBREVIATIONS xvi
CHAPTER
I INTRODUCTION 1
1.1 Introduction 1
1.2 Objectives 2
II LITERATURE REVIEW 3
2.1 Collection and preservation biological evidence 3
2.2 Collection and preservation blood evidence 5
2.3 Forensic DNA analysis 6
2.3.1 Forensic human identification 6
2.3.2 Forensic non-human identification 13
2.3.2.1 Mitochondrial DNA (mtDNA) 14
2.3.2.2 Chloroplast DNA 15
2.3.2.3 Nuclear DNA 15
2.3.3 Steps in forensic DNA analysis 16
2.3.3.1 DNA evidence collection 16
2.3.3.2 DNA extraction from the sample 16
2.3.3.3 Quantification methods in forensic 18
DNA typing
2.3.3.4 Amplified the genetic loci to be examined 19
2.3.3.5 Analysis of amplified DNA 20
2.3.3.6 Allele interpretation 21
2.3.4 Issue in forensic DNA analysis 22
vii
CONTENTS (CONT.)
Page
2.3.4.1 Degraded DNA 22
2.3.4.2 PCR inhibition 22
2.3 Porcine DNA analysis 23
2.4 Artificial DNA degradation methods 24
2.4.1 Sonication 24
2.4.2 UV light 24
2.4.3 DNase enzyme 25
2.4.4 Other methods 25
2.5 Water Quality 26
III MATERIAL AND METHOD 28
3.1 Water types preparation 28
3.2 Conductivity, pH and temperature measurement 29
3.3 Blood sample preparation 29
3.4 DNA extraction by QIAamp DNA mini kit® 29
(Qiagen, Germany)
3.5 DNA quantification 30
3.5.1 UV-VIS spectrophotometer 30
3.5.2 Yield gel 30
3.6 Polymerase chain reaction 30
3.6.1 Primer pairs 30
3.6.2 Thermocycling reaction 31
3.7 Agarose gel electrophoresis 32
IV RESULTS 34
4.1 Measurement of pH and Conductivity of Water Samples 34
4.2 The effect of water samples on untreated bloodstains 35
4.2.1 Amplification of the 211-bp β-actin nuclear 35
DNA fragment
4.2.2 Amplification of the 366-bp β-actin nuclear 36
DNA fragment
viii
CONTENTS (CONT.)
Page
4.3 Testing the effect of carry-over PCR inhibitors in water 38
samples after DNA extraction and purification
4.4 Amplification of the 211-bp β-actin nuclear DNA fragment 42
from swabs of UV-treated bloodstain
4.5 The effect of water samples on UV-treated bloodstain 43
4.5.1 Analysis of 211-bp fragment size of β-actin gene 44
4.5.2 Analysis of 289 bp fragment size of β-actin gene 49
(Primer Pig_F1 and Pig_R2)
4.5.3 Analysis of 289 bp fragment size of β-actin gene 55
(Primer Pig_F2 and Pig_R3)
4.5.4 Analysis of 366 bp fragment size of β-actin gene 61
V DISCUSSIONS 68
VI CONCLUSIONS AND SUGGESTIONS 75
REFERENCES 77
APPENDIX 88
BIOGRAPHY 90