Training Report

on

Basic RECOMBINANT dna technology

Submitted to

Department of Biotechnology

RR Institute of Modern Technology

BKT, Lucknow (U.P.)

Submitted by
Krishnendra dwivedi
B. Tech. (Biotechnology)

Under Supervision of
Er. Madhulika Singh
Head (Division of Biotechnology)

CytoGene Research & Development

Lucknow - 226016
2017
 About CytoGene
CytoGene is an autonomous Registered Organization by Ministry of corporate affairs,
Government of India & Certified by ISO 9001:2008. It provides Training & Research in various
divisions of Life Sciences. It provides both wet & dry Lab, in order to season the students with
the latest and onrushing technological courses, notably oriented to meet the needs of Industry.

We aim at training student with hands on experience at industry level. Our labs are specialized to
provide a halo au courant with hospitals research labs & Bio-Pharma industries.

CytoGene R&D located in Lucknow, one of the growing cynosures in India We embold the
student at Graduate & Post Graduate Level for hands on experience in Projects, Internship & on
Job Training.

On completion of training aspirants are ready to occupy position in various life science firms
with experience candidate in related field.

CytoGene, Lucknow has developed the following Departments.

1) Division of Biotechnology

2) Division of Biochemistry and Enzyme Technology

3) Division of r-DNA Technology &Genetic Engineering

4) Division of Microbiology& Fermentation Technology

5) Division of Immunology& Pathology

6) Division of Molecular Biology

7) Division of Bioinformatics & Genome Analysis

 ACKNOWLEDGEMENT
I express my heartfelt and deep sense of gratitude to Dherandra kumar (HOD), Department of
Biotechnology, RR Institute of Modern Technology BKT- Lucknow for guiding & prompting
me to complete this training.

I would like to give my esteem to Er. Sujeet Kr. Singh (Director) CytoGene Research &
Development, Lucknow for giving me permission to work in this institute.Thereby bestow
platform for students to gain familiarity and working opportunity

I would extend my sincere thanks to Er. Madhulika Singh under whose unvarying supervision
and guidance I was able to complete my entire training.

I would like to thank Mr. Vishal Maurya, Mr. Gyanesh Kumar and Mr. Hemant Kumar
Sharma for their kind lends to hand and insinuation endowed with me during the course of this
training.

At last but not least I would like to thank the Almighty God.

Krishnendra Dwivedi
 DECLARATION
I hereby declare that the training report entitled BASIC RECOMBINANT DNA TECHNOLOGY is
a record of original work carried out by me under the supervision of Er. Madhulika Singh,
CytoGene Research & Development, Lucknow, U.P. (India)-226016.

Date:

Place:

Krishnendra dwivedi
 Content

S.No Title Page No.

1 Chemical Calculation 1
2 Glassware Washing & Sterilization technique 4
3 Ph-meter-its use &calibration 7
4 Preparation of stock solution for DNA isolation 10
5 Primer designing using NCBI Primer-BLAST tool 12
6 Preparation of culture Btoth 18
7 Spread plate Mathod for Culturing and Multiplication 20
8 Plasmid DNA Extraction 22
9 Agarose Gel Electrophoresis 27
10 DNA & RNA quantification by using UV Spectrophotometer 31
11 Polymer chain Reaction 34
12 Restriction Digetion of DNA 39
13 Ligation of DNA EcoR/Hind III fragment in to Puci18 41
14 Preparation of Competent cell and Transformation 43
15 DNA isolation by Metagenomic approach 46
 List of Figures

S. No. Figure Title Page No.

1 5% Chromic acid 8

2 pH meter 9

3 Spread plate 23

4 Streak plate 23

5 Prepared Culture broth(LB broth) 21

6 Agarose Gel Electrophoresis of DNA from soil sample 35

7 Agarose Gel Electrophorsis of Amplified DNA 43

8 Thermo cycler showing the PCR process 43

Restriction digestion sample
9 45

10 Ligated sample 48
Competent cell
11 51

12 Lawn from of Becterial cell growth 51

13 AGAROSE Gel Elecrtophorsis of isolated from soil sample 53

List of Tables
S. No. Table Title Page No.

Table showing the results of Quantification of DNA in UV-Visible

1 41
Spectrophotometer