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LABORATORY ORGANIZATIONS

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CONTENTS�

Research labs overview
Laboratory experiences
Plant tissue culture labs

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RESEARCH LABS

Research Laboratories are workplaces for the conduct of scientific research.

Overview

Building attributes
Lab planning modules
Lab planning concepts
Engineering considerations

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LAB EXPERIENCES

A laboratory is a place where all sorts of scientific and research activities can be performed under controlled conditions
Laboratory experiences can help to enhance national scientific literacy for attaining several educational goals:
Enhancing mastery of subject matter
Developing scientific reasoning
Understanding the complexity and ambiguity of empirical work
Developing practical skills
Understanding of the nature of science
Cultivating interest in science
Developing teamwork abilities

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PLANT TISSUE CULTURE LABS�

Introduction
Basic facilities required in a tissue culture laboratory

Washing area

Glassware/plastic wares

Media preparation area

Transfer area

Culture room

Incubators

Greenhouse

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INTRODUCTION

Plant tissue culture techniques have played pivotal role in the progression of academic and applied plant science.
Initially, this technique was used for studying the role of different hormones.

Now, the major applications are:

Functional characterization of genes
Genetically modified(transgenic)plants
Developing haploid somatic embryos
The large scale production of ornamental plants.
Production of recombinant proteins in liquid culture of plant cells in bioreactors

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CONT…....

Production of novel hybrids by either fusing protoplasts of distantly-related species or embryo-rescue technique.
Molecular, pharmacological and biochemical investigations of plant growth and development such as in vitro flowering.
For the induction of polyploidy, by treating plants with antimitotic agents such

as colchicine.

To produce virus free plants from virus-infected stock, such as potatoes by culturing meristem/tip culture.

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BASIC FACILITIES IN PTC LAB

Plant tissue culture lab is a collection of techniques/methodologies that are employed to grow or maintain plant cells, tissues or organs on a nutrient culture medium under aseptic growth conditions.

Washing area:

Most important place

Used for cleaning and washing of glassware

Consist of sinks

Proper draining boards

Provision for production of deionized water

Specific Space for drying the washed glassware, vessels etc

Storage cabinets for clean & dried material

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GLASSWARE/PLASTIC WARE

Mainly used in:

media preparation & storage
setting up of experiment
tissue culture specific
Pyrex or borosilicate glass vessels are best
Larger volume flasks for preparing culture media
Smaller flasks ,petri plates, test tubes ,jam bottles for growing culture
pre-sterilized disposable polystyrene culture containers(falcon,corning)
Others are beakers, pipette, graduated cylinders

“DETERMINE THE FATE OF EXPERIMENT”

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WASHING OF GLASSWARE

Before using:

Properly wash & autoclave

After experiment:

Immediate cleaning of reusable glassware
Do not allow media to dry
If used for microorganisms….decontaminate and autoclave
Remove all labels and markings
Soak in 5% detergent soln. for min 1 hour
wear facemask and acid resistant apron and gloves
Melt remaining media & discard
Rinse thoroughly with purified water

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MEDIA PREPARATION AREA

Sufficient storage space for:

Chemicals

Culture vessels/glassware

Proper bench space for small equipment e.g:

Stirrers , pH meter, balances, water baths, media dispensing equipment

other necessary equipment includes two autoclaves

one for sterilizing media before initiation

Second for sterilizing waste or left over materials

Refrigerators and freezers to store stock solutions

convection oven for heating the solidified stored medium

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CONT….

careful weighing of all the components
Only analytical grade (AG) chemicals should be used
Follow step-by-step procedure
use purified water instead of tap water
Timely check the functionality of instruments

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TRANSFER AREA

Central place in lab
Should be Well equipped area
Instrument used Laminar flow hood having
Electricity
Ultraviolet light
Compressed air
Vacuum
Positive pressure ventilation unit
High efficiency particular air
During transfer, sterilize periodically with alcohol

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AUTOCLAVE

Aseptic technique for eliminating microbes
Sterile tissues treated with sterile instruments
Media at 15lbs/square inch at 121C for 15 minutes
Disposable sterile plastic ware needs no autoclaving
Filter sterilization for heat liable substances like
Plant growth regulators (auxins, cytokinins)
Antibiotics (ampicillin, kanamycin)

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CULTURE ROOM & INCUBATORS

Another important area
Cultured material need to be incubated in more controlled conditions
Well equipped basic requirements:
Ambient temperature ranges b/w 20 -30
Alarm system to indicate any larger fluctuation
Enough florescent lightening upto 10,000 lux
Both temperature and light shoulb be programmable for 24 hrs
Timers for photoperiodism

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CONT…

Fitted with double doors to kept dust free
Uniform forced -air ventilation
Humidity ranges 20 to 98 % controllable
Air conditioners with regulators managing max & min threshold levels

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GREENHOUSE

Used for plant growing
Obtaining ex-plants