Biotech Lecture 1

1
Biotech Lecture 1

• Lab Safety, Data Record
• Micropipette Usage
• Aseptic technique

2
Safety Regulation and Rules
Every student must know and follow good
laboratory practices in Biotech Center.

• 1) Follow all policies in biotech center.
• 2) The best way to work safely and securely
• 3) What and where safety equipments.
• 4) What to do if an accident occurs.

3
General Safety Procedures

• Never eat, drink, smoke, chew gum in the biotech
  center.
• Cleaning working area after your lab .
• Contaminated supplies are to be disposed of in
  biohazard container.
• After 5 pm, all doors in biotech center should be
  locked even you are still working.
• Check the end of day check-list before leaving.

4
Safety Equipments and Labels

• Safety Equipments
• Goggles must always wear when working with
  chemicals and UV light.
• Masks must always wear when working with
  toxicity chemicals and infectious substances.
• Fire Extinguisher
• Safety Shower and Eye washer
• First Aid Kit

5
Emergency Procedures

• Fire Attack with fire extinguisher and notify
  the instructor, department and wku police
  department.
• Chemical Spills remove all contaminated clothing
  (as appropriate) while using the safety shower.
  For eye contact, Flush with water for 15 minutes.
  Get medical help

6
Data Recording

• On the first page write your name, course, and
  contact information.
• On your note book, you should record
• Date of your experiment.
• Title of your experiment.
• Materials and equipments.
• Experiment procedures.
• Observation and result.

7
Lab Note Format Example

Date 09/26/2005
Experiment
Title E.coli Culture Material and
equipments 5ml of LB medium E.coli colonies in a
LB plate 370 C shaker Procedures
record your procedures Result
sample 1 sample2 amount
15ug 20ug
8
Micropipette
Micropipette 0-20ul Micropipette
0-200ul Micropipette0-1000ul
9
E.coli Bacterial

• Gram-negative, rod-shaped bacterium.
• Easy to grow and amenable to metabolic studies.
• No nuclear membrane
• Chromosome is a large circular duplex molecular.
• Support the replication of DNA plasmids
• Strains used in recombinant DNA work are
  derivatives of K-12
• because their
• restriction modification systems have been
  removed.
• recombination systems are modified to prevent
  rearrangements.
• endonuclease activity has been mutated to
  increase plasmid yields.

10
E.coli Culture - Aseptic Technique
3. Open and sterilize the tops of the tubes
1. Heat the inoculating wire of the loop
2. Pick up 1 colony from the E.coli plate
with the sterilized loop
7. Put the inoculated LB broth tubes to 37 0C
shaker incubator overnight