EDVOTEK® BIOTECHNOLOGY BOOTCAMP
Bacterial Transformation and Transformation Efficiency
TIPS FOR MAKING SOURCE PLATES:
• You can practice this technique by using a marker on the base of the plate and mimicking the steps in Figure 1. • Your students can practice this too! Pass out pieces of paper or white boards and have your students master streaking. • Ensure the agar is fresh and not dried out. Avoid puncturing the plate with the inoculation loop when streaking the bacteria. • Incubate the plate inverted at 37ºC overnight. • Make sure that the BactoBead™ has been stored correctly: Keep the vial capped tightly and in the refrigerator, as exposure to air and moisture will ruin the BactoBead™. • Make sure the BactoBead™ is rehydrated properly with 10 μL recovery broth.
Watch our instructional video here:
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After the incubation:
After the overnight incubation has finished it is important to check on the status of your source plates. The source plate should have isolated colonies that are about 1-1.5 mm in diameter, all the same color, and the agar should not be dried out or hard.
Picking colonies:
When it is time to perform the transformation, students will want to pick up between 5-10 well-isolated colonies for the experiment. To do this, provide the students with sterile inoculation loops or toothpicks and gently swipe the colonies to get them adhered to the loop or toothpick. Then these colonies will be resuspended in ice cold calcium chloride to make them competent! Make sure your students are gentle, and do not pierce the agar with the loop or toothpick.
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