Areej Al-Zahrani
• building 5 – 3rd floor -251
• E-mail : alzahraniareej@yahoo.com
Biotechnology
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Total scores for this lab = 30 marks
Reports 5 marks
Quizzes 5 marks
Performance 3 marks
H.W 2 marks
Final Exam 15 marks
• Final exam DATE :
25-11-2015
Your reference for this lab
my PowerPoint
462 lab not
Biotechnology
• What is the Biotechnology ?
• refers to the use of living organisms or their
products to modify human health and the
human environment.
Biotechnology
lab
• The main objectives of this course are to
introduce the modern and emerging
approaches in Molecular Biotechnology and
its applications in Biochemistry.
microbiological culture
• is a method of multiplying microbial
organisms by letting them reproduce in
predetermined culture media under
controlled laboratory conditions
• All strains of Escherichia coli (E-coli) are
potentially pathogenic
Before the work
• Before opening any culture vessel
• make sure you wear lab coat and gloves . change
your wear after you finish it
• Make sure all you work under the hood
• make sure all the stuff you will need it during the
work , they are near of you
• Think about what kind of cells or bacteria you
with
• one of the important stuff during the work is
alcohol , to clean any thing you contaminate
During the work
• Contamination into cultures can occur from the
air when cultures are opened for manipulation
• so open them for a minimum time
• use smooth movements to reduce sudden air
currents.
• It may also occur from unsterile equipment, as
when a pipette-tip touches your filthy, bacterialaden hands, so be careful with any used pipette,
dispose them immediately.
• make sure you get rid of all the disposal pipet and
medium in right place
• Preventing any organisms or accidental
contaminants from getting out.
• Escape from cultures can occur from dripping
pipettes, from putting used spreaders and
inoculating hoops on the bench without
sterilising them first, by aerosols formed by
blowing bubbles through cultures with
pipettes, or by rapid air movement
(particularly with dry fungal spores).
After the work
• The media and glassware you have been provided will
be autoclaved in steam at 121°C for 20 min. However,
the mouths of culture tubes, inoculating hoops and
spreaders will need to be flamed using with a Bunsen
burner.
• Used pipettes and discarded tubes must be disposed of
into disinfectant.
• Use proper technique whatever the organisms being
used are: you never know when a culture may have
been contaminated, and you owe a duty of care to
yourself and others not to filthy your lab space with
potentially pathogenic bacteria.
Some Helpful hints to remember
when following the methods below.
1. Always wash your hands and spray with 70% ethanol or wear gloves.
2. Always keep the caps on the polystyrene tubes loose so that air can
circulate. Only cap tightly when the cells are no longer growing and are being
stored in the refrigerator until the transformation efficiency has been
calculated.
3. When scraping the frozen cell sample, hold the eppendorf tube at the top
rather than at the bottom, so that the sample does not fully thaw. (one may
want to keep the frozen samples on dry ice.)
4. When doing these experiments always have a negative control. A negative
control is used to show that the media is not contaminated and what is
growing in the media are the bacteria cells and not contaminants.
5. Note that both x-gal and IPTG are light sensitive and therefore should only
be taken out of the light proof box for as brief a time as possible.
6. Ampicillin can cause allergic reactions on contact with skin to those who
are sensitive to penicillin. Do not touch the agar.