genetic engineering is the most successful technique under biotechnology .Here review on various tool of genetic engineering is discussed. #knowmore.

1. VARIOUS TOOLS USED IN GENETIC
ENGINEERING
COMPILED BY
S.AJAY SAMUEL
3rd B.Sc Biotechnology
RVS CAS ,SULUR
COMBATORE

2. INTRODUCTION
• Genetic engineering is a biothenical process of
manipulaion of an organisms gene for the desired
characteristics on the organism which is used for the
study.
• It is a collection of technologies which is used to change
or modify the genetic makeup of the cell which includes
the gene transfer within the species or from other species.
• Here a new DNA molecule is used or an artificially
constructed DNA can be used for the genetic egineering
process

3. CONTINUED.....
• Process involved in the genetic engineering are
1. Gene isolation and cloning
2. Inserting DNA into the host genome
• On this process various tools (chemicals and
machines)are used and they are discussed.

4. POLYMERASE CHAIN REACTION(PCR)
• It is the amplification process of desired DNA molecule
where the amount of DNA is lesser.
• There are five basic reagents used in PCR are template
DNA, PCR primers, dNTPs , PCR buffer and Taq
polymerase.
• Then the amplification process is carried out in a
instrument called THERMOCYCLER were 25-75 cycles of
temprature changes takes place inside the cycler.

5. THERMOCYCLER

6. RESTRICTION ENDONUCLEASE
• Restriction endonuclease/enzymes are also known as
molecular scissors which cuts or digests the DNA
molecule at a specific site for the ligation of desired gene.
• Hundreds of different restriction enzymes, capable of
cutting DNA at a distinct site, have been isolated from
many different strains of bacteria.
• Some enzymes produce sticky end and blunt ends of the
gene
• DNA cut with a restriction enzyme produces many
smaller fragments of varying sizes.

7. EcoRI

8. DNA LIGASE
• DNA is a enzyme which are used for the joining of two
segments or bands of DNA .
• In genetic research, it is often necessary to link two or
more individual strands of DNA, to create a recombinant
strand, or close a circular strand that has been cut with
restriction enzymes. Enzymes called DNA ligases can
create covalent bonds between nucleotide chains. The
enzymes DNA polymerase I and polynucleotide kinase
are also important in this process, for filling in gaps, or
phosphorylating the 5′ ends, respectively.

9. T4 DNA LIGASE

10. POLYMERASES
• The groups of enzymes that catalyze the synthesis of nucleic
acid molecules are collectively referred to as polymerases. It
is customary to use the name of the nucleic acid template on
which the polymerase acts. The three important polymerases
are given below.
• DNA-dependent DNA polymerase that replicates DNA from
DNA.
• RNA-dependent DNA polymerase (reverse transcriptase) that
transcribes DNA from RNA.
• DNA-dependent RNA polymerase that transcribes RNA from
DNA

11. DNA POLYMERASE

12. GEL ELECTROPHORESIS
• Gel electrophoresis is a technique is used for the
visualization of DNA or amplified gene using a phorous
gel method.
• DNA is mixed with the gel loading dye and loaded to a
well formed gel prepared from agarose, where the DNA
band is illuminated by the chemical ethidium bromide.
• Electrophoresis apparatus is used for this process and it
is a migration of DNA using electric charges.

13. ELECTROPHORESIS APPARATUS

14. UV-TRANSILLUMINATOR
• UV-transilluminators are used in molecular biology labs to
view DNA (or RNA) that has been separated by
electrophoresis through an agarose gel. During or
immediately after electrophoresis, the agarose gel is stained
with a fluorescent dye which binds to nucleic acid. Exposing
the stained gel to a UVB light source causes the DNA/dye to
fluoresce and become visible. This technique is used
wherever the researcher needs to be able to view their
sample, for example sizing a PCR product, purifying DNA
segment after a restriction enzyme digest, quantifying DNA or
verifying RNA integrity after extraction.

15. UV-TRANSILLUMINATOR

16. REFERENCE
• explorebiotech.com
• en.wikipedia.org/wiki/Genetic_engineering#Process.
• study.com
• thermofisher.com
• google scholar