State Forensic Science Laboratory

Rajasthan, Jaipur

INTERNSHIP REPORT
 TABLE OF CONTENT

1. Acknowledgement
2. Executive Summary/ Abstract
3. Crime Scene Investigation
• Crime
• Crime Scene
• Steps of Crime Scene Investigation
4. DNA Division
5. Steps of DNA examination
6. After report writing
7. Conclusion
 ACKNOWLEDGEMENT

I would like to express my sincere gratitude to my internship

coordinator Mr.Ramesh Jangid for his great guidelines for internship.

The internship opportunity I had a great chance for learning and

professional development. Therefore, I consider myself as a very lucky
individual and I am also grateful for having a chance to meet so many
wonderful people and professionals who led me though this internship
period.It is a pleasure to thank those who made this internship program
possible. I am heartly thankful to Dr. Ajay Sharma (Director of SFSL,
Jaipur), Dr. Rajesh kumar(Deputy Director, DNA/Sero Division SFSL,
Jaipur), who gave the permission to use all required equipment and
necessary materials to learn various techniques and entire staff for
keeping me in their supervision and giving me exemplary guidance,
constant encouragement to learn and be curious about every single
thing.

Furthermore I would also like to acknowledge with much appreciation to

smt. Bhawna Poonia (Assistant director, DNA division SFSL, Jaipur), Mr.
Ramesh Jangid (SSO, DNA Division SFSL Jaipur) for taking special
consideration for me and taking time out of their busy schedules to
teach me, involve me into cases from opening to analysis part in due
time.
All my lab colleague at the laboratory made it a convivial place to work.
In particular, I would like to thank Mrs. Meenaksh choudhary (JSA, DNA
Division SFSL Jaipur).
Lastly, I offer my regards to all those who supported me in any respect
throughout my internship period.
 Executive Summary
It is a clear fact that the knowledge of construction in Forensic Science
cannot be upgraded without practical experience in each field of the
subject as well as particular subject matter.

This is a report on the internship program that lasted for 20 days form
January 11 to January 31, 2024. I have been learning the vital roles of
Forensic Science. I was able to see and supervise how that they are
multidisciplinary science and technological instruments undertaking
highly specialized scientific work in the service of crime detection, law
and justice. Furthermore, I was working on inward works. I have
developed my theoretical knowledge and practical skills.

This report discuss my overall internship experience that lasted for two
months. This discusses about the DNA/Sero division that determines
Presence of spermatozoa, Presence of human blood and obtain DNA
profile from various types of exhibits such as blood, semen,
bone(marrow), teeth, hair roots, saliva, urine, tissue.

So, I have stated my overall assessment of the whole internship

experience and indicated some improvements that I believe can make it
easy for this program to meet its objective.

In short it explains the experiences I have gained during my stay. And I

have tried to discuss them with much relation to what I have learnt in the
classroom for the last one and half year.
 CRIME
Crime, the intentional commission of an act usually deemed socially
harmful or dangerous and specifically defined, prohibited, and punishable
under criminal law.
For Example: Rape, Murder, Robbery

CRIME SCENE

Any location where crime is committed or suspected to be occurred is

called crime scene.
There are Mainly three types of crime scene are there which are given
below:
1- Indoor Crime Scene: Crime occur inside the door, i.e. robbery in house.
2- Outdoor Crime Scene: Crime occur outside the door, i.e. rape in jungle.
3- Mobile Crime Scene: Crime occur in moving phase.
 CRIME SCENE INVESTIGATION

It is a multidisciplinary process that involves systematic search & meticulous

observation of the crime scene by a dedicated group of forensic investigators
team.

The Seven S'S of Crime-Scene Investigation

 Securing the Scene.

 Separating the Witnesses.

 Scanning the Scene.

 Seeing the Scene.

 Sketching the Scene.

 Searching for Evidence

 Securing and Collecting Evidence.
 DNA DIVISION
DNA Division deals with the cases of violent and civil crimes which comes
under section 302, 303, 376, 5/6 POCSO ACT, SC/ST ACT, etc. Cases like
murder, rape including gang rape, exchange of new born, disputed paternity,
identification of a person in case of bomb blast, terrorist attack, human
trafficking etc are encountered in DNA division.

Physical evidences like Body fluids like blood, semen, saliva, tissues, bones,
teeth, hairs. Saliva present on cigarette-bidi, chewing-gum & gutaka, envelop
and toothbrush etc. Skin, blood and tissue remains in nail scrapings. Foetus,
decomposed, destroyed body parts recovered after bomb blast, fire incident
and exhumed body are examined according to the need.

As soon we open the case, we read the statement of SP carefully, check the
no. Of seals on packet and did examination of the samples mention in the
letter and the samples in which semen is detected.
We get the information of detection of semen by seeing the report of biology.

Steps Of DNA Examination

1. DNA Isolation/extraction.
2. Q-PCR/ Quantification of DNA quality and quantity.
3. Amplification by Polymerage chain reaction(PCR).
4. Genotyping.
5. Report writing.

PRECAUTIONS:
➢ Clean the desk every time before we open new exhibits.
➢ Clean every equipment and machine with the help of alcohol solution.

➢ Wear lab coat, gloves, mask and hair cap to avoid contamination.
➢ Avoid sneezing and coughing on the table where evidences are being
opened.
➢ Protect eyes with close fitting goggles or wrap-around glasses that
contact the skin around the eyes. Ordinary glasses do not provide complete
protection due to gaps.
 DNA Isolation/Extraction

DNA can be extracted from any nucleated cells by automatic DNA

extraction system or Organic extraction method. Isolation process is based
on the principal of magnetic beads. In state forensic sciences laboratory we
use automatic machine methods as per evidence.
Manual methods are: 1- Phenol Chloroform/ Organic Isolation
2- Chelax Method

Automatic Methods are: 1- ABI/ Prepfiler

2- Promega/Maxwell (RSC) 48

The presence of spermatozoa is confirmed by biology department and the

presence of blood stain on exhibit is confirmed by Tetramethylbenzidine
(TMB) Test.
The tetramethylbenzidine test (TMB) is a presumptive chemical test for
identi-fication of blood and is based on the peroxidase like activity of heme
portion of hemoglobin. This test is very sensitive, but not specific. If the
test is positive, confirmatory testing can be performed for conclusive
identification.

REAGENT 1
TMB Reagent:
3,3,5,5‟- tetramethylbenzidine 0.25 gm
Glacial Acetic Acid 25 ml
Store the reagent in a dark colored bottle in refrigerator.

REAGENT 2
3% Hydrogen peroxide (H2O2)
30% H2O2 (SIGMA). 1 ml
Distilled water 9 ml
Mix H2O2 with water and store the reagent in the refrigerator.

Then, the presence of human blood is confirmed by using OBTI.

1.Differential isolation

Used for garments to differentiate male and female fraction. includes

following steps.

1. Sample cutting.
2. Add 400-500μl casework (extraction) buffer + 20μl Proteinase K.
3. Incubation on thermomixer for 4-5 hours at 1000 rpm.
4. Take the tubes and transfer cutting/samples into spin basket followed
by quick spin.
5. Remove spin basket and again spin for 2 minutes.
6. Centrifuge for 2 minutes at 5000 rpm.
7. Seperate
a. Supernatant - as a female part and it's ready for Maxwell.
b. Pellet - Its Male part
I. Washing 2-3 times with 200μl HPLC grade H2O.
II. Add 200μl H2O into pellet tube.
III. Spin/Centrifuge for 50 minutes at 5000 rpm.
IV. Discard the supernatant and again wash with 200μl H2O and spin
at 5000 rpm for 5 minutes.
V. Discard supernatant and in pellet add 100 μl casework buffer +
10μl Proteinase K + 4μl 1- thio glycerol.
VI. Thermomixer for 1 hour at 1000 rpm.

Ready for Maxwell isolation.

2. Maxwell isolation
Used for clothe cutting, tissue, gauze sample

1. Sample cutting.
2. Mixture of - 386μl lysis buffer + 10μl Proteinase K+ 4μl 1- thio glycerol
(400μl) for each sample.
3. Thermomixer for 40 minutes, 56°C +900-1000 rpm.
4. After 40 minutes transfer the cutting into clickfit tube.
5. Spin for 3 minutes at 10,000 rpm.
6. Discard spin basket.
7. Add sample into sample wells + add 200μl lysis buffer
8. Ready for Maxwell isolation.
 3. Bone - tooth extraction
1. sample - Bone and teeth
2. Clean with distill water + Hypo + Alcohol wash.
3. Air dry.
4. Slurry in tissue lyter (add lysis buffer) or Powder in morter pestal.
5. Take 2μl of sample in tube.
6. Bone extraction kit - cocktails -
i. demineralization buffer - 400μl
ii. 1 - thio glycerol. - 10μl
iii. Proteinase K. - 40μl
Add in each sample 450μl

7. Vortex
8. Thermomixer at 56°C + 1000 rpm followed by overnight incubation.
9. After incubation - Quick spin at 10,000 rpm for 3 minutes.
10. Take supernatant and discard debris
11. Add 800μl lysis buffer + 10μl 1 - thio glycerol in each tube.
12. Ready for Maxwell isolation.

For isolation from FTA card

1. Take cutting of FTA card in PCR tubes

2. Add 20μl FTA Buffer in each tube
3. Then run in PCR machine in FTA Isolation program.
4. Take 1μl isolated solution for further Process.
 Start Guide of Maxwell® RCS 48 instrument

.
 QUANTIFICATION - PCR

It is a laboratory technique for quantitation or to measure the amount of

reagents used while doing PCR. It also detect male chromosome and we
can determine the dilution percentage. It works on the principle of
thermocycler.

PROCEDURE:
1. Sample (1μl)+ Reaction mix (5μl) + Primer (4μl).
2. Centrifuge for 4-5 minutes at 10,000 - 20,000 rpm.
3. Load the sample in RT-PCR machine for 40 minutes
4. After that see the value or the graph between cycle threshold or
quantity for further processing in PCR.
5. If the value of large autosomal is small that means DNA is present in
small quantity so no dilution is done but if the value of large autosomal
is large then dilution is done in PCR with the help of nuclease free
water.

Fig. – QuantStudio 5 by Thermo Fisher Scientific

 POLYMERASE CHAIN REACTION :
PCR machine is used to amplify selected polymorphic loci called short
tendem repeat (STR) of isolated DNA.
PCR is widely used method for making millions of copy of a specific DNA
sample to make our study easier.

The full reaction is of 25μl.

It has three kits: 1- Global -Filer

2- Y- Filer
3- Fusion- mix

They all have same components and procedure the only difference in the
quantity of primer and reaction mix. It is patent to specific company.

Global-Filer: Master mix (3.75μl) and Primer Mix (1.25μl), after that we add
water and sample for dilution as per our need to complete the reaction.

Y-Filer: Reaction mix (5μl) and Master Mix(2.5μl) after that we add water and
sample for dilution as per our need to complete the reaction.

Fusion: Primer mix (1.25μl) and Reaction Mix(2.25μl) after that we add water
and sample for dilution as per our need to complete the reaction.

Fig. – Thermocycler by Ependorf

GENOTYPING :
It works on the principle of Capillary Electrophoresis. We do
sequencing to obtain Electropherogram. Sequencer tray has 96 blocks
and 24 sample can run at time per slot. 1block has 8 column and 3
column can run at a time.

Gene Mapping IDX software is used to see the results.

Capillary of Sequencing machine is fragile in nature and made up of

silica. System has POP (Performance Optimize Polymer) which should
be change after 386 runs. 4μl POP is used per sample.

REQUIREMENTS:
ILS (Internal Lane Standard)
HIDI (Highly Deionized Formamide)
Sample
Allelic LADDER

We use 8.5μl HIDI for the stability of denatured DNA so that they
remain single standard and 0.5μl ILS for the position and size of
markers in EPG. LADDER is used after 23 blocks to provide
standardization.
Before performing genotyping we centrifuge through plate centrifuge
and denaturation through PCR to make it Ss- DNA from Ds- DNA
because genetic analyzer reads Ss-DNA sequence.

Fig. – 3500 XL Genetic Analyzer by ABI

REPORT WRITING:
Report writing is one of the most important step of DNA profiling. The report
of DNA profiling is admissible in court of justice under section 293 CrPc .
The report must be consize and laymen type.

The report writing may be separated in three categories :

• Basic detail of the case
• Result of DNA analysis
• Conclusion

Basic detail :- In this, FSL fills all the necessary details of the case like FIR
No., case no., section, Date, Police Station, District, etc.

Result :- The Results section will contain the results of all analyses including
screening re-sults, DNA results, comparisons/conclusions, result Data and
statistics. The results section may list results in a bulleted format or a
narrative format. All items submitted for Forensic Biology & DNA
examinations will be addressed in the Report. Items not examined will be
addressed in the Report as not having been examined. Results of tested
alleles of examined samples should be in tabular form in the Report. Based
on results of examination, observations or interpretations should be drawn.

Conclusion :- -Conclusion will follow the RESULTS section. This section is

reserved for reports containing multiple items tested and hence more results
to be reported. It may be incorporated into the report to assist in streamlining
results, thus creating a more reader friendly report. When incorporated, it will
contain results of comparisons, result data, and statistics. For example-

 Whether it’s matched or not.

 Biological mother or not.

 Biological father or not.

 Biological relative or not.

Steps after report writing:

1. The evidence has to be sealed by covering thread and knot.

2. The evidence sealed by using divisional seal which has to be printed

properly.

3. The evidence bag has to be send to OGE Section for dispatch with
report and H form.

4. The packed bag include all details such as case details (FIR No., Date,
Police Station, District, Examiner’s signature, packet mark, exhibit
mark).
 CONCLUSION

I hereby conclude my report by saying that this was my first internship and it
was really a great experience in terms of learning and fun as well. From my
training at DNA division I was able to get a better understanding of scientific
approach of every task. It was arollercoaster ride while working in divisions
and enjoyed working and looking forward to gain my knowledge by
consistent efforts in future. This internship experience was important in
improving my academic knowledge and to get more insight of a practical.
Thank you so much for granting me such wonderful experience as well
guidance.
I got to see many astounding things such as weapons, bone, and many other
unique evidences which are not seen on regular basis. The helpful and
supported staff of SFSL made this experience to remember.
The feel of handling real crime cases is starling & super motivating to
become a forensic expert & solving the mystery frenetic to reduce the crime
rate of country.