Exam 1

Question 1

What is Molecular Biotechnology?

Answer 1

Science discipline that is based on the ability of researchers to transfer specific units of genetic information from one organism to another

Question 2

What was the first commercial product using recombinant DNA technologies?

Answer 2

Human Insulin

Question 3

What is recombinant DNA technology?

Answer 3

insertion of a gene into a DNA vector (often a plasmid) to form a new DNA molecule that can be perpetuated in a host cell

Question 4

DNA Insert

Answer 4

extract DNA from a donor organism

Question 5

cloning vector

Answer 5

extracted DNA is joined to another DNA entity

Question 6

mutagenic plasmid

Answer 6

the cloning vector insert DNA construct

Question 7

transformation

Answer 7

introduction of DNA into a bacterial host strain

Question 8

types of restriction endonuclease

Answer 8

type I
type II
type III
type IV

Question 9

which type of restriction endonuclease is the most common

Answer 9

Type II, often called restriction enzymes

Question 10

why do we use type II restriction enzymes

Answer 10

they recognize a defined, symmetric sequence
cuts within the recognition site

Question 11

common problems when performing a restriction digestion

Answer 11

incomplete digestion
incomplete dephosphorylation of cloning vector
problems with ligation

Question 12

gene SOEing

Answer 12

gene Splicing by Overlap Extension - a sequence independent method for site-directed mutagenesis and/or recombination of DNA molecules. it is based on the idea that a PCR product can be engineered by adding or changing sequences at its ends so that the product can itself be used to prime DNA synthesis in a subsequent overlap-extension reaction to create mutant or recombinant molecules.

Question 13

Types of Genetic Transformation

Answer 13

Electroporation
heat shock
conjugation

Question 14

electroporation

Answer 14

the action or process of inducing DNA or chromosomes into bacterial or other cells by using a pulse of electricity to briefly open the pores in the cell membranes

Question 15

Conjugation

Answer 15

the transfer of genetic material between bacterial cells by direct cell-to-cell contact or by a bridge like connection between 2 cells

it is one of the mechanisms of horizontal gene transfer

Question 16

heat shock

Answer 16

uses a calcium rich environment provided by calcium chloride to counteract the electrostatic repulsion between the plasmid DNA and bacterial cellular membrane

Question 17

Steps of PCR

Answer 17

Denaturation
Annealing
Extension

Question 18

Why do you want to use shorter templates in PCR

Answer 18

the less template used, the easier and faster the amplification process is done, allowing the yield to grow exponentially

Question 19

Northern

Answer 19

RNA

Question 20

Southern

Answer 20

DNA

Question 21

Western

Answer 21

Proteins

Question 22

Molecular Hybridization

Answer 22

complimentary pieces of DNA and RNA can go back together

Question 23

Reverse Transcriptase

Answer 23

running the central dogma backwards

Question 24

chemical synthesis of DNA

Answer 24

machine that automates the chemical reactions of DNA synthesis have made the production of single stranded oligonucleotides (<50 nucleotides) routine procedure

Question 25

Phosphoramidite Steps

Answer 25

washing
detitrylation
washing
activation and coupling
washing
capping
oxidation
back to start to add next nucleotide
end the cycle

Question 26

coupling efficiency

Answer 26

determined by spectrometric ally monitoring released trityl groups; should be greater than 98% at each step

Question 27

bioton

Answer 27

streptavidin - used to pull stuff out of a system

Question 28

DNA sequencing

Answer 28

use of technologies to determine the identity and order of the 4 nucleotide bases in a segment of DNA