Detection of IS6110 Gene by Real Time PCR in

Mycobacterium Tuberculosis Bacteria

Training Report
Submitted to:

Graphic Era University, Dehradun, Uttarakhand, India

For partial fulfillment of award of degree in bachelor of technology in

biotechnology

Under the guidance of:

Supervisor
Co-supervisor

Dr. Narotam Sharma

Dr. Nita Garg
Scientist, Central Molecular Research Laboratory,
Professor and Head,

Biochemistry Department
Biochemistry Department

Center for Molecular Research Laboratory, SGRRIMHS, Department

of Biochemistry, Patel Nagar, Dehradun, Uttarakhand, India
 Submitted by:

Bhavya Mudgal B.Tech Biotechnology ( 7th semester)

DECLARATION
I, undersigned hereby declare that the project entitled, Detection of IS6110
by RT PCR in Mycobacterium Tuberculosis Bacteria, is a study conducted at
Center for Molecular Research Laboratory, SGRRIMHS, Department
of Biochemistry, Patel Nagar, Dehradun, Uttarakhand, India under the
supervision of by Dr. Narotam Sharma, Scientist, Central Molecular Research
Laboratory, Shri Guru Ram Rai Institute of Medical & Health Sciences, Patel
Nagar, Dehradun, Uttarakhand, India . The pragmatic finding in this project is
based on the data and information collected by myself.

Bhavya Mudgal

B.Tech Biotechnology(7th Semester)

 ACKNOWLEDGEMENTS

It is my privilege to express our sincerest regards to our Head of the

Department Dr. Ashish Thapliyal, Graphic Era University, Dehradun. Dr.
Narotam Sharma, Scientist at Central Molecular Research Laboratory of
Shri Guru Ram Rai Institute of Health and Medical Sciences, Dehradun.

I am also thankful to my project coordinator Dr. Nita Garg for their valuable
inputs and able guidance, encouragement, whole hearted cooperation and
constructive criticism throughout the duration of my project.

I deeply express my sincere thanks to Dr. Narotam Sharma for

encouraging and allowing us to present the project on the topic Detection
of IS6110 Gene by Real Time PCR in Mycobacterium Tuberculosis
Bacteria and I am also thankful to Mr. Satish Chandra Nautiyal who
continuously helping me regarding the project and give me the chance to
work on the diagnostic samples that helpful for me in the future.

I am also thankful to all the staff members of Biochemistry Department of

Shri Guru Ram Rai Institute of Medical & Health Sciences for helping me

whenever I approached them.I cannot express in words the sacrifices of my

parents. Their motivation, inspiration and care are incredible. I will always

remain indebted to them

.
 CONTENTS
S.N PARTICULARS PAGES
O
1. INTRODUCTION 1
2. AIMS AND OBJECTIVES 3
3. MATERIALS AND METHODS 5
4. RESULT 19
5. CONCLUSION 22
6. REFERENCES 23
 List of Figures

S.N Ligands to figures Page

o. No.
1. Mycobacterium tuberculosis 2
2. Shri Guru Ram Rai Institute of Medical and Health Sciences 6
3. Biosafety Level-3 Laboratory at Central Molecular Research Laboratory 7
4. Reagent Storage Area 9
5. Reagent Preparation Area 9
6. BSL-1 9
7. BSL 1 Spinwin Centrifuge 10
8. Dress Changing Area 10
9. Enter in BSL-2 10
10. Biosafety Laminar -2 10
11. BSL-2 Centrifuge 10
12. Real Time PCR 10
13. Area-2: Bio Safety Cabinet 12
14. Rotor Gene Q 15
15. Polymerase Chain Reaction (Denaturation, Annealing, Extension & 18
Storage)
16. Pie Chart 20