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Lab Final Study Guide

Terms in this set (70)

Enzyme
proteins that speed up chemical reactions without creating heat or becoming part of the product
Substrate
...
Active site
...
Product
...
Activation energy
...
Cellobiase
...
PNG
...
p-nitrophenol
...
How does a spectrophotometer work?
...
How to read a standard curve relating p-nitrophenol and absorbance
...
How to determine reaction rate of cellobiase enzyme
...
Plasmid
...
pGLO
...
GFP
...
bla
...
Beta-lactamase
...
Ampicillin
...
araC
...
Arabinose
...
Central dogma
instruction from DNA are used to make a functional product (proteins).
Difference between -DNA and +DNA tube
...
Calculate μg of pGLO used
...
Calculate transformation efficiency
...
Restriction endonuclease
...
Restriction sites
...
Restriction fragment length polymorphism
...
Ethidium bromide
...
Loading buffer
...
How does gel electrophoresis work?
...
Which side of the gel box do the wells of the gel go?
...
Which direction do the DNA fragments move?
...
How to read gel electrophoresis
...
How to match band pattern to a plasmid cut into X pieces
...
Convert between μL and mL
...
T/F
GFP is naturally found in bacteria
False
T/F
Bacteria naturally have plasmids
False
Why do we need CaCl2 when transforming bacteria?
Help facilitate transformation process
T/F
DNA is positively charged
False
If the black end of the box is negative and the red end of the box is positive, which end will the wells of the gel in?
Black bc its negative
Which of the following genes does pGLO NOT have?
A. bla
B. araC
C. GFP
D. Beta-lactamase
Beta-lactamase
If a plasmid has 4 restriction sites and is digested at all sites, how many bands will it have if run on a gel?
4
What color is DNA when doing a gel?
Colorless
What is the term for the process of taking RNA and turning it into a protein?
Translation
Translation
...
Transcription
...
If a bacteria is transformed with the pGLO plasmid and plated on an LB plate, will it grow?
yes
If a bacteria is transformed with the pGLO plasmid and plated on an LB plate, will it glow?
no
Cellobiase converts ____ into ____.
PNG; p-nitrophenol
What portion of the linear equation derived from the enzyme graphs indicates the rate of reaction? y=mx+b
m
araC requires what molecule to activate and allow translation of GFP protein?
arabinose
If bacteria is transformed with pGLO and plated on LB+amp+arabinose, will it grow?
yes
If bacteria is transformed with pGLO and plated on LB+amp+arabinose, will it glow?
yes
List 3 factors that affect enzyme activity.
1. Enzymatic inhibitor
2. pH
3. Temperature
What color is p-nitrophenol?
Yellow
Where did we obtain the enzyme for lab 9?
mushrooms
What is the purpose of creating a standard curve during the first session of lab 9?
To have a baseline of enzyme activity to compare to the enzyme activity in the experiment where a variable is changed
You used a micropipette to transfer CaCl2 and LB broth. Convert 200 μL (microliters) to ___ mL.
2.00 x 10^-1
Which plates were expected to grow single colonies in the bacterial transformation lab?
(+) pGLO LB/Amp
(-) pGLO LB/Amp
(+) pGLO LB/Amp/Ara
(-) pGLO LB
(+) pGLO LB/Amp
(+) pGLO LB/Amp/Ara
(-) pGLO LB
Which plates were expected to glow in the bacterial transformation lab?
(+) pGLO LB/Amp
(-) pGLO LB/Amp
(+) pGLO LB/Amp/Ara
(-) pGLO LB
(+) pGLO LB/Amp/Ara
What would happen to bacteria transformed with pGLO ​[(+) pGLO] and grown on plates with the antibiotic tetracycline (very different than ampicillin)? Will the transformed bacteria grow or not at all?
Bacteria will not grow
In your own words, what is the central dogma and how does it relate to lab 9?
The central dogma is: instruction from DNA are used to make a functional product (proteins).

The plasmid uses GFP gene to create pGLO enzyme which allows the bacteria to glow

GFP= the instructions
pGLO= functional protein from GFP1.
GFP stands for _____.
Green Fluorescent Protein
pGLO is a ___.
plasmid
T/F
Bacteria transformed with pGLO should grow in the presence of ampicillin and only fluoresce green under ultraviolet light when arabinose is added.
True
What gene does a plasmid need in order to allow bacteria to grow in the presence of ampicillin antibiotics? What protein does this gene encode for?
bla gene, beta-lactamase protein
After the pGLO plasmid is cut with HindIII endonuclease, how many DNA fragments (pieces) should we end up with? How many bands should we end up with on the gel?
2;2
If you DNA has pGLO the in the electrophoresis chamber there will be ___ lines
2
The smaller the fragment, the ____ distance the fragment will migrate; the larger the fragment, the ____ distance of migration.
Larger; shorter
What is the name of the endonuclease used in lab 11?
Hindlll
T/F
The wells of the gel (containing your DNA) are placed near the negative end of the gel electrophoresis box.
True
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