Talha Nasir

22100260
Bio lab section 3
Assignment 1
 Media preparation
Introduction:
lysogeny broth (LB MEDIA) is used for the growth of certain aero torelent bacterial species such
as E. coli, Bacillus subtilius, Staphylococcus auerus or Staphylococcus epidermidis and different
yeasts including Saccharomyces and Candida species.

Objective of the day:

our objective was to create LB Broth and LB agar to be used in a medium to grow bacteria

Materials used and their reasons:

Electronic mass balance to measure weight of substances
Plastic spoon to pour powdered substances into balance
Piece of paper as a container to measure weight of materials

the following ingredients were used to create 100 ml of LB-BROTH and LB-Agar:

around 100 ml of water

1g bacto-tryptone
0.5g yeast extract
1g NACL

Method:
for lb-broth, add the following to a jar: 1g bacto-tryptone ,0.5g yeast extract, 1g NACL. After
this add distilled water to make the solution 100ml and then sterilize it by autoclaving.
for lb-agar, add the same ingredients but with the addition of 1.5g agar gel.
this media will be used to grow bacteria after autoclaving is complete.

Observations:
Before auto claving was done, the lb-broth was a green-yellow liquid and lb-media was a light
yellow liquid
 Agarose gel electrophoresis

Introduction:
Agarose gel electrophoresis is the easiest method to separate and analyze DNA

Objective of the day:

To prepare agarose gel and use it for electrophoresis

Materials used:
Buffer chamber and gel tray for location of electrophoresis to be conducted
Tape for ends of casting tray
Power source and leads for electrophoresis to be conducted
Agarose is used in making agarose gel
Comb is used to make wells
10x TAE used as a buffer
Loading dye

Method:
Firstly add 1.5g of agarose in a jar by using an electronic balance, then add 50 ml of 10* TAE
using a measuring cylinder. Heat this solution using a microwave to melt the agarose
(and make solution colorless) then cool it down and add it to the tray. Place the comb to
make wells (to be used in loading of gel). Place the gel tray in the electrophoresis tank.
Add the buffer to the buffer chamber and use a p-20 pippete to add 5 x10^-6 liters of
loading gel into each well, do this fast to avoid spillage of gel and also avoid touching
the wall surrounding the gel to avoid the gel going back in the pipette. After this attach
the power supply and the process starts.

Observations:
Bromophebol Blue travelled less far than Xylene Cyanol
 PIPETTE
(used pipette)
Introduction:
A pipette is a laboratory tool used to transfer a measured volume of liquid from one
container to another

Objective of the day:

To learn how to efficiently use pipettes

Materials used and their reasons:

p-20 pipette used for volume of ranges 5 to 20 microliters
p-100 pipette used for volume of ranges 10 to 100 microliters
p-1060 pipette used for volume of ranges 10 to 100 microliters
unique flasks for each pipette to attach to.
A particular liquid to use the pipette for

Method:
Attach the flask to a pipette. Press the button and insert the flask into the liquid to be collected
and release the button to collect the liquid. Press the primary button to release the liquid and
double press if it isn’t released correctly. Press the secondary button eject the flask
 Contamination

Introduction:
Transient flora are microorganisms present on our skin for short periods of time that may
represent a threat to a person’s health.

Objective of the day:

To compare transient flora on thumb before and after washing

Materials used and their reasons:

Nutrient agar plates to grow bacteria

Method:
Divide the plate into two equal sides, label one as washed and the other as unwashed, touch
the unwashed side by your thumb and cover the petri dish immediately. Wash your hands using
hand wash and tap water and dry them, rub them with ethanol to further sterilize them. After this
touch the washed side with your thumb and cover the petri dish again. Store the petri dish at 37
degrees for at least 18 hours for growth to occur.

Observations:
Many different results were obtained throughout the group:
A few results as expected resulted in initially having very little to heavy growth initially
and a significant reduction after washing. This means that hands were cleaned
thoroughly
Some results resulted in little to no difference or an increase in growth after washing.
This means that either hand washing was not done properly, or hands were re-
contaminated after washing by atmosphere, or by closing the tap, or by opening the
petri dish or some other reason.