Genetics and Biotechnology

Question 1

What is the difference between the proteome and functional proteome?

Answer 1

The proteome is the entire set of proteins encoded by the genome. The functional proteome is the complete set of proteins currently expressed.

Question 2

How can proteins present in muscle be evaluated?

Answer 2

SDS-PAGE electrophoresis

Question 3

What is SDS and what is it used to do?

Answer 3

It is a strong anionic detergent that provides a negative charge that can be used to linearize proteins.

Question 4

What is the overall basis for how SDS-PAGE electrophoresis?

Answer 4

1. Proteins are denatured in SDS
2. A negative charge is added to the proteins
3. The proteins are loaded onto polyacrylamide gels
4. The proteins are separated based on molecular weight

Question 5

Following SDS PAGE electrophoresis, intensity of the bands depends on what?

Answer 5

protein concentration

Question 6

Myofibrillar proteins are extracted in _____ salt concentrations. Sarcoplasmic proteins are extracted in ____ salt concentrations.

Answer 6

• high

- low

Question 7

On SDS PAGE gel, where are heavier proteins found?

Answer 7

At the top

Question 8

What two changes does SDS impart on the proteins that help it run on the gel?

Answer 8

SDS denatures the proteins and applies a negative charge

Question 9

What kinds of fibers are in skeletal muscles?

Answer 9

Both red and white fibers

Question 10

What are the four methods of isolating DNA?

Answer 10

DNAse, RNAse, ethanol precipitation, and phenol/chloroform

Question 11

How do you determine DNA purity and concentration?

Answer 11

Agarose Gel Electrophoresis

Question 12

What technique is used to visualize DNA fragments on agarose gel?

Answer 12

Southern Blot

Question 13

Describe the agarose gel electrophoresis method.

Answer 13

DNA is cut up using restriction enzymes and agarose gel electrophoresis, which separates DNA fragments according to their size, allows you to visualize the fragments.

Question 14

The equipment that is used for DNA fragment viewing. The DNA is placed in the chamber, and electric current is run, and the DNA migrates toward the positive end.

Answer 14

Gel apparatus

Question 15

What are the three steps in DNA isolation?

Answer 15

Extraction, purification, electrophoresis

Question 16

What does PCR allow you to do?

Answer 16

Make millions of copies of the DNA without a need for the host cell

Question 17

What are the basic steps of PCR?

Answer 17

1. Denature- break apart the DNA
2. Annealing- anneal primers to flank DNA targets
3. Extension- extend primers using taq DNA polymerase and the dNTPs

Question 18

A _____ ____ is used to amplify DNA via PCR.

Answer 18

Thermal Cycler

Question 19

What are the types of PCR?

Answer 19

Reverse Transcriptase PCR and qPCR or “Real Time” PCR

Question 20

When the sample for PCR is in the thermal cycler, what is happening?

Answer 20

Heats up to denature the double stranded DNA, cools to annealing temperature to allow the forward and reverse primers to bind, heating occurs to allow extension of the primers

Question 21

WHat should be on the gel to evaluate an agarose gel with PCR?

Answer 21

A ladder, PCR product, positive/negative, and housekeeping gene