lab background

Question 1

what is tranformation

Answer 1

Bacteria take in DNA from the environment

Question 2

conjugation

Answer 2

Bacteria share DNA by touching eachother

Question 3

plasmids

Answer 3

small DNA circles with extra genes

Question 4

What do plasmids carry

Answer 4

genes for things like antibiotic resistance.

Question 5

Why do scientists use DNA transfer in the lab?

Answer 5

to make bacteria gain new traits.

Question 6

what is recombinant DNA

Answer 6

dna made by combining genes from different sources.

Question 7

Plasmid vector

Answer 7

a plasmid used to carry new genes into bacteria

Question 8

selectable marker

Answer 8

a gene that lets scientists find bacteria with the plasmid

Question 9

what is an inducible promoter

Answer 9

a switch that turns a gene on only when a specific molecule is present

Question 10

what molecule turns on a GFP in the lab?

Answer 10

IPTG

Question 11

GFP

Answer 11

green fluorescent protein that glows under blue light

Question 12

How do scientists purify GFP

Answer 12

column chromatography

Question 13

How do scientists check the size and purity of GFP

Answer 13

SDS-PAGE

Question 14

why do we centrifuge the cells?

Answer 14

to pellet the bacteria at the bottom of the tube and work with the concentrated cells

Question 15

Why do we add ice cold CaCI2 to the cells?

Answer 15

to make the cells competent, Calcium ions help the cell membrane take in DNA

Question 16

why do we incubate the cells on ice?

Answer 16

to keep the cells stable and help them become competent

Question 17

What is the Competent Cell Solution (CCS) used for?

Answer 17

to resuspend the cells before transformation keeps the cells ready and stable for DNA uptake.

Question 18

Why do we heat-shock the cells?

Answer 18

helps DNA enter the bacteria by creating a temperature difference.

Question 19

Why do we add recovery broth?

Answer 19

to help bacteria recover after heat shock, cells repair their membranes and start expressing plasmid genes.

Question 20

why do we use -DNA and +DNA plates

Answer 20

-DNA is control (no plasmid) +DNA is transformed, to compare growth and confirm transformation worked.

Question 21

how do we visualize GFP

Answer 21

Use UV light. GFP glows green, showing which bacteria were transformed.

Question 22

Transformation efficiency

Answer 22

number of transformed cells perNumber of transformed cells per μg of DNA. measure our success in the experiment.

Question 23

What factors affect transformation efficiency?

Answer 23

Cell health, pipetting technique, incubation times, and plasmid quality.

Question 24

How can plasmids be inserted into bacteria?

Answer 24

Through bacterial transformation.

Question 25

What happens to bacteria after taking up a plasmid?

Answer 25

They can express new traits from the plasmid genes, like antibiotic resistance or GFP.

Question 26

Can you outline the steps of bacterial transformation?

Answer 26

1. Make cells competent (CaCl2/CCS, cold incubation) 2. Add plasmid DNA (+DNA tube) 3. Heat shock to help DNA enter 4. Allow recovery in nutrient broth 5. Plate on selective media 6. Observe growth and traits (e.g., GFP)

Question 27

What can you demonstrate in the lab with transformation?

Answer 27

Inserting plasmid DNA into bacteria and observing new traits.

Question 28

how was diabetes diagnosed in the 1800s

Answer 28

doctors tasted urine for sweetness

Question 29

how was diabetes treated in the 1800s

Answer 29

diets and herbal remedies

Question 30

when was insulin discovered

Answer 30

1921

Question 31

how is diabetes diagnosed today

Answer 31

blood glucose tests and continuous glucose monitors

Question 32

How is diabetes treated today

Answer 32

insulin, meds, diet, exercise, and monitoring devices.

Question 33

diabetes

Answer 33

body cant properly control blood sugar

Question 34

genetic engineering

Answer 34

changing dna to give organisms new traits

Question 35

insulin

Answer 35

hormone that lowers blood sugar by helping cells absorb glucose

Question 36

iptg

Answer 36

chemical that turns on genes like GFP in the lab

Question 37

Antibiotic Sensitivity

Answer 37

When bacteria are killed or stopped by an antibiotic.

Question 38

Antibiotic Resistance

Answer 38

When bacteria survive in the presence of an antibiotic.

Question 39

Promoter

Answer 39

: DNA sequence that starts transcription of a gene.

Question 40

Multiple Cloning Site

Answer 40

DNA region with many cut sites for inserting genes.

Question 41

Origin (Ori)

Answer 41

DNA sequence that allows plasmid replication.

Question 42

Selectable Marker

Answer 42

Gene that lets scientists identify bacteria with the plasmid.

Question 43

Protein Purification

Answer 43

Protein Purification

Question 44

Chromatography

Answer 44

Method to separate molecules by size or charge.

Question 45

Column Chromatography

Answer 45

Using a column with beads/matrix to separate proteins.

Question 46

Native Protein

Answer 46

Protein in its natural 3D shape, still functional.

Question 47

SDS-PAGE

Answer 47

Denaturing gel method; separates proteins by size only.

Question 48

Coomassie Blue Stain

Answer 48

Dye used to see proteins in a gel.