PCR PCR is a method widely used in Molecular biology to make many copies of a specific DNA segment. Using PCR it is possible to generate thousand millions of copies of a particular section of DNA from very small amount of DNA. PCR was originally developed in 1983 by the American Biochemists Kary Mullis. He was awarded the Nobel Prize in Chemistry in 1993 for his pioneering work.

1. Govt. Institute Of Forensic
Science, Nagpur
Presented By,
Palash A. Mehar
B.Sc.(Forensic Science) Part-III

2. Polymerase Chain Reaction(PCR)
eppendorf -
Mastercycler® pro S
Topic:-

3. Polymerase Chain Reaction (PCR):-
 PCR is a method widely used in Molecular
biology to make many copies of a specific
DNA segment.
 Using PCR it is possible to generate
thousand millions of copies of a particular
section of DNA from very small amount of
DNA.
 PCR was originally developed in 1983 by
the American Biochemists Kary Mullis. He
was awarded the Nobel Prize in Chemistry
in 1993 for his pioneering work.
Kary Mullis
(Invented PCR in 1983)

4. Materials Required For PCR:-
The following components are needed to perform PCR in
the laboratory,
 DNA template
 A heat-stable DNA polymerase (like Taq Polymerase)
 Two short, single-stranded DNA molecules that serve as
primers
 All four nucleotide triphosphates
 Buffers
 Thin walled tubes
 Thermal cycler (a device that can change temperatures
dramatically in a very short period of time)

5. Steps involved in PCR:-

6. 68 Billion
copies at
the end of
35 cycle

7. Intended use of
Mastercycler® pro
• Used for temperature control of aqueous solutions, suspensions or
emulsions in closed micro test tubes for enzymatic reactions, typically
the polymerase chain reaction (PCR).
• Exclusively intended for indoor use.
• All country-specific safety requirements for operating electrical
equipment in laboratories must be observed.
• The product can be used for training, routine and research
laboratories in the areas of life sciences, industry or chemistry. This
product is intended to be used for research purposes only.

8. Product Description
Master variants
The mastercycler pro is available
in three variants,
• Mastercycler pro
• Mastercycler pro S
• Mastercycler pro 384
Front and rear view,
1) Lid handle For opening, closing, locking and
unlocking the heated lid.
2) Heated lid
3) Heating plate
4) Thermoblock
5) Ethernet port
6) Mains connection socket with power switch
0 = switched off , I = switched on.
7) Eco and Term switches
8) CA N out port
9) CA N in port
10) Control panel See detailed drawing (see p.
20)
11) Flap USB port cover.
12) Name plate

9. Control Panel:-
1) stop key Stop the running
program.
2) Standby key To switch the
device to power save mode.
3) Display
4) Number pad
5) Status lamp In the case of
devices with a thermoblock with
64+32 wells, the status lamp in
the control panel is deactivated.
The active status lamp for
devices with a thermoblock with
64+32 wells is located on the
heated lid.
6) del key To delete marked objects
and characters to the right of the
cursor.
7) enter key To confirm entries and
open selection lists.
8) next key Move the cursor to the
next input field.
9) Arrow keys Move the cursor.
10) Softkeys Function depends on
the software dialog and is
indicated in the display above the
softkey.
11) start key Start the selected
program.

10. Featutres of mastercycler pro :-
All variants are provided with the following features:
1) Triple Circuit Technology
All Mastercycler pro thermomodules feature extremely fast heating and cooling
rates. Triple Circuit Technology ensures a homogeneous temperature distribution and
enables the generation of specific temperature gradients for PCR optimizations.
2) SteadySlope
SteadySlope Technology ensures that the heating and cooling ramp rates for thermal
modules are identical in both gradient and normal operation. That means you can expect
identical temperature control characteristics in both optimization and routine experiments,
as well as a reliable transfer of the optimization results to the routine application.
3) vapo.protect
All variants of the Mastercycler pro have a vapo.protect heated lid. A flexible
pressure element ensures that force is applied evenly to the surface of the tubes or plate.
This reduces evaporation from the reaction sample effectively. The vapo.protect heated lid
performs automatic tube height adjustment for all sample tubes and PCR plates.

11. Contd...
4) TSP
The heated lid of the Mastercycler pro series is equipped with
TSP technology (Thermal Sample Protection). The thermoblock is
continuously kept at 20°C during the lid heating phase. This means TSP
reduces the thermal load on your samples and minimizes the probability
of non-specific product formation during the PCR.
5) Self-test function
You can use the integrated self test function of the
Mastercycler pro in combination with the control panel with USB ports
to check whether the thermoblock is functioning properly without the
need for any further tools. A separately available USB key is required for
the self test function.
6) Control
The Mastercycler pro is controlled via the control panel. It can
also be controlled via the PC software CycleManager pro

12. Contd...
• Highest temperature control speed due to silver block: up
to 6°C/s.
• Standard PCR possible in under 30 min.
• Pulse function for device-driven hot start for avoiding
non-specific PCR product formation.
• Freely programmable temperature gradient over 12 rows.
• Extended gradient range of up to 24°C.
• Holds up to 96 PCR tubes (0.2 ml) or a 96-well PCR
plate.

13. Structure of Program Editor:-
1) Number of the program step
2) Block temperature [°C]
3) Hold time [mm:ss]
4) Average temperature for gradient
steps
5) Program step highlighted marking
6) Step number within the cycle
7) Active input field
8) Extended program step
Marking of gradient steps
9) Number of cycles

14. Operation of mastercycler ,
1) Login
2) Creating folders and programs
3) Copying folders and programs
4) Editing programs –
 Select program
 Open program editor
 Press Header function key
 Set lid functions
 Confirm input
 Press insert function key
 Select the program step
 Enter temperature, hold times, cycle number.
 Select program step
 Press the option function key
 Set the gradient, increment, and ramp
 Confirm input.
 Press the save function key
 Press exit function key

15. Operation of mastercycler ,
5. Stopping and starting program
• Mark the program
• Insert the sample tubes(max.
Sample volume 100 μL per single
reaction)
• Close the heater lid
• Start the program
• Confirm the input
6. You can use the status view to
carry out following program
commands,
 Stop
 Abort
 Resume

16. Status View