Lab 1- Lab Notebook, Isolation Plate, Subculturing, Types of Media

Lab Notebook- Goal

The goal is to allow someone to read over the notebook and recreate the experiment.

Lab Notebook-Key Elements

This includes:

• Table of Contents

• Clear Experimental Procedures/Data

  • Use ink

• Sign and Date Each Entry

• Do Not Tear Out a Page

Lab Notebook-Setting Up

The table of contents (first 2 pages) have the title of lab, page number, and date. Each experiment needs the title, date, procedure, and results. Initial the bottom corner when done with a page.

Isolation Plate- Setting Up Plate

1. Draw a ‘T’ for the quadrants. Number the quadrants, write the date, initial, organism written on the back.

2. Flame the loop to sterilize.

3. Cool loop on media plate and streak the bacteria.

4. Flame loop and then transfer some bacteria from the first to second quadrant. Streak again.

5. Flame loop and then transfer some bacteria from the second to third quadrant. Streak again.

6. Flame loop once more for sterilization and put back.

Isolation Plate- Day 4

The first quadrant will have confluent growth, quadrant two is more diluted, and quadrant three has isolated colonies.

• Confluent growth essentially looks like grass or rows of bacteria. It is of high density.

• Isolated colonies have discrete, single dot growth. it

Subculturing

• This is transferring cells to a new vessel to maintain the culture.

  • The slant has confluent growth.

• Isolated colonies have a pure culture meaning they can be used to subculture.

• Agar slants give the bacteria a greater surface area for the tube to grow.

Procedure

1. Sterilize the loop and pick up bacteria from one of the tubes and tap it on the agar. Repeat this with the other bacteria; tap it in the same spot.

2. Sterilize and spread.

3. On day 4, subculture each of the bacteria and incubate.

4. On day 5, transfer it to the fridge.

Types of Media

1. Enriched: This has nutrients that fastidious and pathogenic bacteria need to grow.

2. Selective: This has salts and dyes to stop unwanted microbes from growing, allowing the organism to be isolated. INHIBITORS

3. Differential: This has pH dyes and blood to reveal the differences in metabolism. This will cause the colonies to change colors.