Research on Genetics and Replication in Biology is the Basis for Experiment

LAB REPORT
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Background information
Research on genetics and replication in Biology is the basis for this experiment
References
The lab manual; pages E-1 to E-7
Purpose
The purpose of this experiment is to identify an ideal strain for transformation by using enzymes. The blue/white essay will be used to identify the strain currently housed by the plasmid needed to transform. The objective is to identify the bacterial strain that can be transformed to create an organism that makes high concentrations of fetal hemoglobin, which can be used to treat sickle cell anemia. The promoter upstream of the LacZ gene can be induced by lactose to produce protein at high levels. Various strains labeled A, D, L, Y will be assessed.
Experiment
Materials
16 culture tubes with blue caps, bacterial culture, 37° incubator, ice bucket, 1M NaC03, Spec20 and cuvettes, 16 culture tubes with teal caps, sugars for induction (20% glucose), toluene, ONPG, 28°C water bath, timer.
* Label the 16 culture tubes: label the first 1-8
* Obtain cultures from the incubator
* Add 5mls of the wild type E. coli strain culture to tubes 1-4
* Add 5mls of culture with the mutant E Coli to tubes 5-8, labeled: A, D, Y, L
* Add water to each tube as necessary so that the final volume of each tube is 5.42mls
* Return all the cultures to the shaking incubator for 45 minutes. During this time, the cells will translate mRNAs and translate proteins.
* Before the 45mins incubation ends, label 16 blue cap tubes as before 1-8 and add two drops of toluene to each tube. Cap the tubes immediately and place them on ice
* At the end of the incubation, add 2mls of the appropriate bacterial culture to each tube. Vortex and return the tubes to ice.
* After collecting all samples, place the tubes in a 28°C water bath for 30 minutes swirling occasionally. During this time, the cells will weaken, allowing them to take up the substrate.
* Transfer all your tubes to the lab bench and add 400mls of ONPG solution to each tube in intervals of 15 seco...