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Isolation of Streptomycin Resistant Mutant - Lab Experiment | BI 203, Lab Reports of Microbiology

Material Type: Lab; Professor: Lester; Class: MICROBIOLOGY; Subject: Biological Sciences; University: Montgomery College; Term: Unknown 1989;

Typology: Lab Reports

Pre 2010

Uploaded on 03/11/2009

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Microbiology Lab Experiment Changes
Experiment #: none
Title: Gradient Plate: Isolation of Streptomycin Resistant Mutant
Live Organisms: E. coli
Changes: Procedure
(Work in pairs or if in groups, do two sets per group)
1. TSA deeps have been melted and are being held at 45-50°C in a
water bath.
2. Follow diagrams below. Pour one melted agar deep into an
empty Petri dish, then put a colored pencil (it has the right thickness)
under one end and wait 15 minutes.
3. Next remove pencil from under Petri dish. Add 1.0 mL of colored
Streptomycin solution to the remaining melted agar deep. Invert tube
to mix and pour on top of first layer of agar. The streptomycin
solution contains a colored dye to allow the gradient to be visualized.
Wait 15 more minutes.
4. After the agar plate has completely solidified, spread 0.2 mL of
the E. coli culture onto the surface of the agar. Your instructor will
demonstrate this technique.
Take Home Lesson: Why would you expect to find any streptomycin resistant mutants?
What is the purpose of the streptomycin? Does streptomycin induce (cause) mutations? What
are some of the possible mechanisms a mutant might use to circumvent the streptomycin?
First agar deep
Put pencil under one end. Wait
15 minutes for agar to solidify.
Second agar deep +
1.0 mL streptomycin Wait 15 minutes then spread 2
drops of E. coli on surface.

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Microbiology Lab Experiment Changes

Experiment #: none

Title: Gradient Plate: Isolation of Streptomycin Resistant Mutant

Live Organisms: E. coli

Changes: Procedure (Work in pairs or if in groups, do two sets per group)

  1. TSA deeps have been melted and are being held at 45-50°C in a water bath.
  2. Follow diagrams below. Pour one melted agar deep into an empty Petri dish, then put a colored pencil (it has the right thickness) under one end and wait 15 minutes.
  3. Next remove pencil from under Petri dish. Add 1.0 mL of colored Streptomycin solution to the remaining melted agar deep. Invert tube to mix and pour on top of first layer of agar. The streptomycin solution contains a colored dye to allow the gradient to be visualized. Wait 15 more minutes.
  4. After the agar plate has completely solidified, spread 0.2 mL of the E. coli culture onto the surface of the agar. Your instructor will demonstrate this technique.

Take Home Lesson: Why would you expect to find any streptomycin resistant mutants?

What is the purpose of the streptomycin? Does streptomycin induce (cause) mutations? What are some of the possible mechanisms a mutant might use to circumvent the streptomycin?

First agar deep Put pencil under one end. Wait 15 minutes for agar to solidify.

Second agar deep + 1.0 mL streptomycin

Wait 15 minutes then spread 2 drops of E. coli on surface.