Term
| Is the Cathode positive or negative? |
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Definition
|
|
Term
| Is the Anode positive or negative? |
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Definition
|
|
Term
| Is the cation positive or negative? |
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Definition
|
|
Term
| Is the anion positive or negative? |
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Definition
|
|
Term
| What is the principle of electrophoresis? |
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Definition
| Migration of charged particles in an electric field |
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Term
| Do the Anions migrate towards the Cathode or the Anode? |
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Definition
|
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Term
| Do the Cations migrate towards the Cathode or the Anode? |
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Definition
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Term
| Name 3 Types of DNA (RNA) electrophoresis methods |
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Definition
Agarose Electrophoresis
Polyacrylamide Gel Electrophoresis
Capillary Electrophoresis |
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|
Term
True or False:
Agarose Electrophoresis - Migration of DNA is Independent of Charge |
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Definition
|
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Term
True or False:
Agarose Electrophoresis - Migration of DNA is Independent of Size |
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Definition
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Term
| Agarose Electrophoresis - What pH range is DNA negatively charged? |
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Definition
|
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Term
| Why is DNA negatively charged at pH 7-8? |
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Definition
|
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Term
| Is the Mass to Ratio consistent with DNA in agarose electrophoresis? |
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Definition
| Yes. 1 Phosphate group/nucleotide |
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Term
| Agarose Electrophoresis - Separation is based on _____ of nucleic acid |
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Definition
|
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Term
| Which size fragments migrate faster? |
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Definition
|
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Term
| Which size fragments migrate slower? |
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Definition
|
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Term
| What are the Pros of Agarose Electrophoresis? |
|
Definition
Large range of separation (200 bp - 50,000 bp)
Easy to set up & perform
Non-Toxic |
|
|
Term
| What is the Con of Agarose Electrophoresis? |
|
Definition
| Lower resolving power (sizes <100 bp hard to see) |
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Term
| What type of Electrophoresis is most commonly used in Clincial Lab? |
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Definition
|
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Term
| What is the most common concentration of the agarose gel? Which does UAMS MLS use? |
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Definition
|
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Term
| The Higher the concentration, the More Dense the matrix, and the Smaller the Pores, the ______ DNA fragments migrate |
|
Definition
|
|
Term
| Do you use a higher concentration or a lower concentration of agarose gel for small fragments? |
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Definition
|
|
Term
| Do you use a higher concentration or a lower concentration of agarose gel for large fragments? |
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Definition
|
|
Term
| What do Buffers do in Agarose Electrophoresis? |
|
Definition
Establish pH
Provide ions for current conductivity |
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|
Term
| If you use only water in Agarose Electrophoresis what happens? |
|
Definition
No ions in gel
Conductivity lost
DNA won't migrate |
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|
Term
| If the buffer is not diluted properly in Agarose Electrophoresis what can occur? |
|
Definition
Ion concentration too high
Too much heat may be generated
Gel will melt
DNA won't migrate |
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|
Term
| What is the content of the Loading Buffer for Agarose Electrophoresis? |
|
Definition
| 10-20% glycerol & tracking dyes |
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|
Term
| What does glycerol do to the sample? |
|
Definition
Increases sample density
(Sample "drops to bottom of well" when loading gel) |
|
|
Term
| What is the purpose of tracking dyes? |
|
Definition
| Help visualize samples for loading and for tracking migration of samples through gel |
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|
Term
| What is the most common dye for DNA electrophoresis? |
|
Definition
|
|
Term
| What does the fluorescent Ethidium bromide dye do between bases of Nucleic Acids? |
|
Definition
|
|
Term
| Is Ethidium bromide a mutagen? |
|
Definition
| Yes: Probably a carcinogen as well |
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|
Term
| Should you wear gloves to protect your hands from being stained when you use Fast blast DNA stain? |
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Definition
|
|
Term
| Is Fast Blast DNA Stain toxic? |
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Definition
|
|
Term
| Is Fast Blast DNA Stain carcinogenic? |
|
Definition
|
|
Term
| What is Polyacrylamide gel Electrophoresis (PAGE)? |
|
Definition
| Cross-linked polyacrylamide polymer |
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|
Term
True or False:
The pores in PAGE are more controlled and uniform |
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Definition
|
|
Term
| Does PAGE have very high resolution or very low resolution? |
|
Definition
|
|
Term
True or False:
PAGE can distinguish fragments with 1 bp difference |
|
Definition
|
|
Term
| PAGE is good for ____ fragments |
|
Definition
|
|
Term
| What range is PAGE good for? |
|
Definition
|
|
Term
| Is PAGE a new method for DNA sequencing? |
|
Definition
| No. It is the traditional method. |
|
|
Term
| What are the 3 disadvantages of PAGE? |
|
Definition
Neurotoxin
Need large gel for DNA use
Must be poured between glass plates |
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|
Term
| Which type of electrophoresis uses very small-bore, long capillary tubes? |
|
Definition
| Capillary electrophoresis |
|
|
Term
| Which type of electrophoresis is replacing PAGE? |
|
Definition
| Capillary electrophoresis |
|
|
Term
| What does PCR do to its target? |
|
Definition
|
|
Term
| Does PCR give a qualitative or quantitative result? |
|
Definition
|
|
Term
| What are 2 of the uses of RFLP and REs? |
|
Definition
Identity Testing
Studying microorganisms involved in outbreaks |
|
|
Term
Put the following procedure in order for RFLP/RE:
*Separate RFLPs with electrophoresis
*Treat DNA with one or more RE
*Measure distance unknown fragments migrate, use graph to determine size of unknown
*Measure distance
*Include standard DNA ladder in 1 lane
*Graph results
*Determine size |
|
Definition
*Treat DNA with one or more RE
*Separate RFLPs with electrophoresis
*Include standard DNA ladder in 1 lane
*Measure distance
*Graph results
*Measure distance unknown fragments migrate, use graph to determine size of unknown
*Determine size |
|
|
Term
| What is Pulse-Field Gel Electrophoresis used for? |
|
Definition
| To separate large DNA fragments (large RFLPs) |
|
|
Term
True or False:
PFGE - get better separation if precisely change direction of electrical current |
|
Definition
|
|
Term
| What is the Goal of PulseNet? |
|
Definition
| Rapidly detect outbreaks and take preventive action |
|
|
Term
| PulseNet - what is the organism treated with? |
|
Definition
|
|
Term
|
Definition
| Network of local, state, and federal agencies |
|
|
Term
True or False:
PulseNet - Food-borne disease-causing bacteria is tested |
|
Definition
|
|
Term
True or False:
The following organisms are examples of bacteria tested with PulseNet:
E. coli O157:H7
Salmonella
Shigella
Listeria
Campylobacter
Vibrio cholerae |
|
Definition
|
|
Term
True or False:
The pattern of the Organisms is entered into the PulseNet data base |
|
Definition
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