ACKNOWLEDGEMENT In the completion of the project work, I am overwhelmed by a secured feeling of satisfaction and achievement.

In these prime moments of elating pleasure I take this opportunity to offer my sincere regards to all those who made this training period an immemorial one. I express my sincere gratitude to Prof. Nuzhat Husain (Dean, HOD, Dept. of Pathology, RMLIMS) for providing me an opportunity to undertake my internship at Dr. Ram Manohar Lohia Institute of Medical Sciences, Lucknow. I am thankful to Mr. Shivbrat Upadhyay for his support, cooperation and motivation provided to me during the internship and for continuous inspiration, presence and blessings. I also extend my sincere appreciation to Dr. P. K. Paul (HOD, Amity Institute of Biotechnology) and Dr. Tuhin Rashmi (Faculty Guide) who provided her valuable suggestions and precious time in helping me to accomplish my project report. Lastly I would like to thank my pillars of life i.e. my parents and friends who provided me with the basic things which I required and showed full confidence in me and gave me full moral support.

AVIJIT ZUTSHI DATE: PLACE: NOIDA

DECLARATION I, Avijit Zutshi, student of B. Tech Biotechnology, studying in 7th semester, hereby declare that the Dissertation report on Factor V Leiden Mutation in cases with Hemophilia A submitted to Amity Institute of Biotechnology, in the partial fulfilment of degree of Bachelor of Technology in Biotechnology is the original work conducted by me at Dr.Ram Manohar Lohia Institute Of Medical Sciences, Lucknow. The information and data given in the report is authentic to the best of my knowledge. This internship report is not being submitted to any other place for award of any other degree, diploma and fellowship. The publications, if any, coming out of the Project work shall be done only after the consent of the Company/Industry (if any) where a part of / whole project work has been carried out. Any indiscretion arising out of my work or conduct shall be my responsibility.

AVIJIT ZUTSHI DATE: PLACE: NOIDA

CONTENTS

S.No.

Topic

Page No. 13 4 11

1.

Introduction

2.

Review of Literature

3.

Aims and Objectives

12

4.

Materials and Methods

13 14 23 24 25 26 40 41 43

5.

Instrumentation

6.

Reagents Used

7.

Experimental Techniques

8.

Result

9.

Discussion and Conclusion

44 45 46

10.

Appendices

11.

References

47

ABBREVIATIONS

APS APC Bp DVT dNTP EDTA EtBr FVIII FV PAGE PCR RFLP SNP TAE TBE TEMED

Ammonium Per Sulphate Activated Protein C Base Pair Deep Vein Thrombosis Deoxyribo-nucleotriphosphate Ethlenediaminetetraacetic acid Ethidium Bromide Factor VIII Factor V Polyacrylamide Gel Electrophoresis Polymerase Chain Reaction Restriction Fragment Length Polymorphism Single Nucleotide Polymorphism Tris-Acetate-EDTA Tris-Borate-EDTA N, N, N, N-tetramethylethylenediamine