Duong_Richard_Prelab5 - Richard Duong MCRO 251 Lab 28...

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Richard Duong MCRO 251 Lab 28 September 2010 Week 5 Pre-Lab Objective: This lab has two parts, each with a different objective. The first objective is to examine bacteria transformation, a technique where competent bacteria take up DNA and either incorporates it into their genome or forms a plasmid. Insertional activation will also be observed in this experiment using different plasmid vectors resulting in differences in phenotype. The second objective is to examine the effects that microorganisms have on your hands based on the uses of different sanitizers. Procedure: 1. Check streak plates from last week to see if isolated colonies were obtained. 2. Calculate the plaque forming units per milliliter (PFU/ml) of the original sample based on a set of two plates. a. Use the formula Stock concentration (PFU/ml) = # plaque/volume plated (ml) x 1/concentration plated. 3. Report to lab instructor so that an average of the class can be calculated. Bacterial Transformation 1. Mark tube containing competent E. coli as to identify it as yours. a. Keep this tube over the majority of experiment (they are very fragile). 2. Add 10 micro liters of DNA prep from “D” tube to competent cells on ice using a micropipette. 3. Close the lid and gently mix the DNA and bacteria. 4.
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Duong_Richard_Prelab5 - Richard Duong MCRO 251 Lab 28...

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