Cloning The Clone

Front 1

Outline the process of cloning

Back 1

DNA extraction
Isolating the gene of interest
Transformation
Culturing
Radioactive probing

Front 2

DNA Extraction

Back 2

Plasmid removed from bacterial cell
Chromosomal DNA removed from human cell containing gene of interest

Front 3

Isolating the gene of interest

Back 3

Plasmid and chromosomal DNA cut w/restriction enzymes
Stick ends; unpaired bases, generated

Chromsomal DNA and bacterial plasmid mixed

Sticky ends complementary base pair with DNA ligase
Recombinant DNA formed

Front 4

Transformation

Back 4

Recombinant DNA inserted into bacterial cell via transformation

Only some bacterial cells will have gene of interest

Front 5

Culturing

Back 5

Bacteria cultured in nutrient medium -> binary fission
exponential growth replicates plasmid and clones gene of interest
Bacterial cells streaked on petri dish, multiply overnight

Front 6

Radioactive probing

Back 6

Bacterial colonies probed for gene of interest with complementary radioactively labelled DNA probe or antibodies

Front 7

Transgenesis

Back 7

May occur after cloning process

Electoporation/micropartical gun for plants

Fine Glass Needle: animals

Front 8

Which step is Transformation?

Back 8

STEP 3

Front 9

Which step is DNA Extraction?

Back 9

STEP 1

Front 10

Which step is culturing?

Back 10

STEP 4

Front 11

Which step is isolating the gene of interest?

Back 11

STEP 2

Front 12

Which step is radioactive probing?

Back 12

STEP 5