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23 Cards in this Set

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Bacterial Transformation?
process where bacterial cells take up naked DNA molecules
When will the DNA replicate the foreign DNA along with their own DNA?
if the foreign DNA has an origin of replication recognized by the host cell DNA polymerases
Competent?
bacteria that are able to uptake DNA
What are the 3 ways bacteria can be put into DNA?
conjugation, transuction, transformation
Conjugation?
DNA is transferred from one bacterial donor cell to recipient cell via sex pilli
Transduction?
DNA is introduced into bacteria by injection from a virus
Transformation?
bacterial cells take up naked DNA molecules
How does ampicillin prevent bacteria from growing?
prevent bacteria from making a cell wall
Why would you want to insert DNA into bacteria? 2
-to make more DNA
-to make more protein
What will be inserted into E.Coli for this lab?
pGLO plasmid
What genes do pGLO contain?
bla, GFP, araC
Role of bla gene?
provides resistance to the antibiotic ampicillin by using beta lactamase protein
role of GFP gene?
codes for green fluorescent protein
Role of araC gene?
promotes the production of GFP by producing araC protein
How are competent bacteria made?
treatment with calcium chloride followed by heat treatment at 42 degrees celsius
Role of cold temperature in transformation?
helps the DNA adhere to cell membrane of E.Coli
Role of high temperatures in transformation?
increases the permeability of the cell membrane to DNA
What is used to help the bacteria recover from heat shock?
briefly incubate bacteria with non-selective growith media (LB)
Transformation efficiency?
quantative value that describes how effective you were at getting a plasmid into bacteria
-represents the number of transformed colonies per microgram of DNA added
Transformation effiency formula?
total number of cells growing on plate/amount of DNA spread on agar
Formula to determine the amount of DNA spread on the agar(microg)?
total amount of DNA(microg) *fraction of DNA used
Formula to determine fraction of DNA used?
volume spread on plate(micoL)/total sample volume in test tube (microL)
Determine amount of pGLO in DNA(microg)?
concentration of DNA (microg/microL) * volume of DNA (microL)