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Page 1: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

DNA Technologies

Page 2: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Biotechnology

Dolly and surrogate Mom

Genetically modified rice.

Embryonic stem cells and gene therapy

Page 3: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Biotechnology

Biotechnology, defined broadly, is the engineering of organisms for useful purposes.

Often, biotechnology involves the creation of hybrid genes and their introduction into organisms in which some or all of the gene is not normally present.

Fourteen month-old genetically engineered (“biotech”) salmon (left) and standard salmon (right).

Page 4: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

DN

A

Tech

nolo

gy

DNA technology is the chemical manipulation of the genotypes and resulting phenotypes of organisms such that living organisms are modified

Alternatively, no-longer-living organisms or their no-longer-living parts may be analyzed chemically at the level of genotype

DNA technology has revolutionized how scientists study the genetics, biochemistry, even the ecology and evolutionary biology of organisms

Genetic engineering is the artificial manipulation of the genetic material of organisms, including the creation of novel genetic material (i.e., novel nucleotide sequences)

Biotechnology is the development of novel biological products, indeed whole industries are now devoted to the production and analysis of biological materials

Page 5: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Biotechnology

We’ll examine:

Animal cloning

Gene cloning for pharmaceutical production

The promise and perhaps perils of embryonic stem cells

DNA fingerprinting

Genetically modified foods and the American-European opinion divide.

Page 6: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Animal Cloning

Dolly and her surrogate mother.

Page 7: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Why Clone Animals?

To answer questions of basic biology

Five genetically identical cloned pigs.

For herd improvement. To satisfy our desires (e.g. pet cloning).

For pharmaceutical production.

Page 8: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Is Animal Cloning Ethical?

The first cloned horse and her surrogate mother/genetic twin.

As with many important questions, the answer is beyond the scope of science.

Page 9: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

USU’s Contribution – A Cloned Mule and the First Cloned Equine

Page 10: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

The Biotechnology of Reproductive Cloning

Even under the best of circumstances, the current technology of cloning is very inefficient.

Cloning provides the most direct demonstration that all cells of an individual share a common genetic blueprint.

Page 11: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Saved by Cloning?

Some are firm believers while many view these approaches to be more of a stunt.

Note the use of a closely related species, a domestic goat, as egg donor and surrogate mother.

Page 12: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

(Science (2002) 295:1443)

Carbon Copy– the First Cloned Pet

Significantly, Carbon Copy is not a phenotypic carbon copy of the animal she was cloned from.

Page 13: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

The Next Step?

Highly unlikely.

Attempts at human cloning are viewed very unfavorably in the scientific community.

Page 14: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

So how do we do all of this???

Page 15: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Recombinant DNA methods Restriction enzymes

Enzymes from bacteria Used to cut DNA molecules in specific places Enable researchers to cut DNA into

manageable segments Vector -molecule carrier of DNA fragment into

cell Transformation: uptake of foreign DNA into cells

Page 16: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Cutting DNA with a restriction enzyme

Page 17: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Rest

rict

ion

E

nzy

mes

Page 18: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Restriction Endonucleases

• A Restriction Endonucleases will cut both strands of a DNA duplex at a specific place

• These “places” need not be directly opposite:

• Note that the above enzyme is EcoRI, the first restriction endonuclease characterized

5’…GAATTC…3’ 3’…CTTAAG…5’

5’…GAATTC…3’ 3’…CTTAAG…5’

5’…G -OH P-AATTC…3’ 3’…CTTAA -P HO-G…5’

5’…G -OH P-AATTC…3’ 3’…CTTAA -P HO-G…5’

Page 19: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Sti

cky

En

ds

Page 20: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Most R.E. RecognitionSequences are Palindromes

G^AATT-CC-TTAA^G

G^GATC-CC-CTAG^G

A^GATC-CT-CTAG^G

GC^GGCC-GCCG-CCGG^CG

Nodeba Bob Abedon

Page 21: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

More

RE

E

nzy

mes

Enzyme Sequence Product

EcoRI G^AATTC 5’ sticky ends

BamHI G^GATCC 5’ sticky ends

Bg1II A^GATCT 5’ sticky ends

PvuI CGATC^G 3’ sticky ends

PvuII CAG^CTG Blunt end

MboI G^ATC 5’ sticky ends

HindIII A^AGCTT 5’ sticky ends

HinfI G^ANTC 5’ sticky ends

Sau3A G^ATC 5’ sticky ends

AluI AG^CT Blunt end

TaqI T^CGA 5’ sticky ends

HaeIII G^GCC 5’ sticky ends

NofI GC^GGCCGC 5’ sticky ends

Page 22: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Restriction Enzymes are Enzymes That Cut DNA Only at Particular Sequences

The enzyme EcoRI cutting DNA at its recognition sequence

Different restriction enzymes have different recognition sequences.

This makes it possible to create a wide variety of different gene fragments.

Restriction enzyme animation

Page 23: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Recombinant DNA, Gene Cloning, and Pharmaceutical Production

DNA can be cut at specific sequences using restriction enzymes.

This creates DNA fragments useful for gene cloning.

These are mature and widely utilized biotechnologies.

Page 24: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

DN

A

Lig

ase

Upon ligation we now have recombinant

DNA

Page 25: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Lig

ati

on

Page 26: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Splicing foreign DNA into a vector Foreign DNA and plasmid DNA cut with same restriction

enzyme Produces linear molecules with complementary single-

stranded ends Recombinant DNA created by mixing so sticky ends pair DNA ligase forms covalent bonds, linking the two fragments

Page 27: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

DNAs Cut by a Restriction Enzyme Can be Joined Together in New Ways

These are recombinant DNAs and they often are made of DNAs from different organisms.

Page 28: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Plasmids

Page 29: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Plasmids are Used to Replicate a Recombinant DNA

Plasmids are small circles of DNA found in bacteria.

Plasmids replicate independently of the bacterial chromosome.

Replication often produces 50-100 copies of a recombinant plasmid in each cell.

Pieces of foreign DNA can be added within a plasmid to create a recombinant plasmid.

Page 30: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Tra

nsf

orm

ati

on

Note that plasmid is vector that

carries DNA into recipient

cells via transformation

Other vectors include viruses (transduction)

as well as otherwise inert

projectiles

Page 31: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Harnessing the Power of Recombinant DNA Technology – Human Insulin Production by Bacteria

Page 32: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

http://www.sumanasinc.com/webcontent/anisamples/molecularbiology/plasmidcloning_fla.html

animation

Page 33: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Human Insulin Production by Bacteria

6) join the plasmid and human fragment

and cut with a restriction enzyme

Page 34: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Human Insulin Production by Bacteria

Mix the recombinant plasmid with bacteria.

Screening bacterial cells to learn which contain the human insulin gene is the hard part.

Page 35: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Route to the Production by Bacteria of Human Insulin

A fermentor used to grow recombinant bacteria.

This is the step when gene cloning takes place.

The single recombinant plasmid replicates within a cell.

Then the single cell with many recombinant plasmids produces trillions of like cells with recombinant plasmid – and the human insulin gene.

One cell with the recombinant plasmid

Page 36: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Route to the Production by Bacteria of Human Insulin

The final steps are to collect the bacteria, break open the cells, and purify the insulin protein expressed from the recombinant human insulin gene.

Page 37: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Route to the Production by Bacteria of Human Insulin

Overview of gene cloning.

Cloning animation

Page 38: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Pharming

These goats contain the human gene for a clot-dissolving protein that is produced in their milk.

Pharming is the production of pharmaceuticals in animals engineered to contain a foreign, drug-producing gene.

Page 39: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Genetically Modified Foods

Many of our crops in the US are genetically modified.

Should they be?

Page 40: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

GM Crops are Here Today

Source: Pew Initiative on Food and Biotechnology, August 2004.

Page 41: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Methods for Plant Genetic Engineering are Well-Developed and Similar to Those for Animals

Page 42: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Golden Rice is Modified to be Provide a Dietary Source of Vitamin A

Worldwide, 7% of children suffer vitamin A deficiency, many of them living in regions in which rice is a staple of the diet.

Golden rice (yellow) with standard rice (white).

Page 43: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Genetically Modified Crops

Genetically Modified Cotton (contains a bacterial gene for pest resistance)

Standard Cotton

Page 44: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

GMOs, Especially Outside the US, Are a Divisive Issue

Protesters at the 2000 Montreal World Trade Summit

European sentiment

Page 45: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Current Concerns by Scientists Focus on Environmental, Not Health, Effects of GM Crops

The jury’s still out on the magnitude of GM crop’s ecological impact, but the question is debated seriously.

Page 46: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Current Concerns by Scientists Focus on Environmental, Not Health, Effects of GM Crops

Page 47: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Genetic probes Segments of single-stranded DNA that can hybridize to

complementary base sequences in target gene Usually a radioactive piece that “labels” the DNA Southern blot technique

Page 48: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Using a geneticprobe to find bacterial cellswith a specificrecombinant DNA molecule

Page 49: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Amplifying DNA in vitro by PCR Small amount of double-stranded DNA DNA precursors Specific nucleic acid primers Taq DNA polymerase

DNA is denatured Primers attach to primer-binding site on each DNA strand Each strand acts as template for DNA synthesis

Page 50: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Poly

mera

se C

hain

R

eact

ion

Cloning allows the amplification

of genotype (DNA) as well as

phenotype (proteins)

If all you really need is the DNA, then PCR is an

easy way to amplify DNA

without cloning

Page 51: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Amplification of DNA by PCR

Page 52: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

PC

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PC

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Page 54: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Gel Electrophoresis

Page 55: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Loading Gel

Page 56: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Load

ed

G

el

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Ru

n

Gel

Page 58: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

DNA, the Law, and Many Other Applications – The Technology of DNA Fingerprinting

A DNA fingerprint used in a murder case.

What are we looking at? How was it produced?

The defendant stated that the blood on his clothing was his.

Page 59: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

DNA Fingerprinting Basics

Different individuals carry different alleles.

Most alleles useful for DNA fingerprinting differ on the basis of the number of repetitive DNA sequences they contain.

Page 60: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

DNA Fingerprinting Basics

If DNA is cut with a restriction enzyme that recognizes sites on either side of the region that varies, DNA fragments of different sizes will be produced.

A DNA fingerprint is made by analyzing the sizes of DNA fragments produced from a number of different sites in the genome that vary in length.

The more common the length variation at a particular site and the greater the number the sites analyzed, the more informative the fingerprint.

Page 61: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

A Site With Three Alleles Useful for DNA Fingerprinting

DNA fragments of different size will be produced by a restriction enzyme that cuts at the points shown by the arrows.

Page 62: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

The DNA Fragments Are Separated on the Basis of Size

The technique is gel electrophoresis.

The pattern of DNA bands is compared between each sample loaded on the gel.

Gel electrophoresis animation

Page 63: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Possible Patterns for a Single “Gene” With Three Alleles

In a standard DNA fingerprint, about a dozen sites are analyzed, with each site having many possible alleles.

Page 64: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

A DNA Fingerprint

When many genes are analyzed, each with many different alleles, the chance that two patterns match by coincidence is vanishingly small.

DNA detective animation

HGP fingerprinting page

Page 65: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

DNA and the Law

SLT 3/8/05

Some applications of DNA fingerprinting in the justice system.

Page 66: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

2-D

Pro

tein

E

lect

rop

hore

sis

Page 67: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

DNA typing

Page 68: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Sou

thern

B

lott

ing

Page 69: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Northern Blot

A Northern Blot is the same as a Southern Blot-except instead of probing for DNA- the prob is for RNA

Used to detect whether or not transcription is occuring

Page 70: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Western Blot

Same as Southern/Northern except the probe is for protein

Used to detect whether a particular gene is being “turned on” or “off”

Page 71: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

DNA sequencing Based on chain termination method Yields information about

Structure of gene Probable amino acid sequences of its encoded proteins

Page 72: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Chain termination method of DNA sequencing

Page 73: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

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Page 78: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Transgenic organisms Foreign DNA incorporated into their genetic material Gene target

Single gene inactivated or “knocked out” “Knockout mice” used in studying genes

Mutagenesis screening Male mice treated with mutagens and studied

Page 79: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

A transgenicmouse

Mouse on rightis normal; mouseon left is transgenicanimal expressingrat growth hormone

Page 80: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Transgenicrice

“Golden rice”shown intermixedwith white ricecontain highconcentrationsof beta-carotene

Page 81: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Safety guidelines Safety concerns

Introduction of transgenic organisms into the environment

Health effects on humans from consuming GM crops Safety measures

Special facilities designed to hold pathogenic organisms

Science of risk assessment

Page 82: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Gen

e

Th

era

py

One example of this might

be done… is essentially cloning into

animals

Page 83: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Injecting DNA into Egg

Page 84: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Genome Comparisons

Page 85: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

The Stem Cell Concept

A stem cell is an undifferentiated, dividing cell that gives rise to a daughter cell like itself and a daughter cell that becomes a specialized cell type.

Page 86: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Stem Cells are Found in the Adult, but the Most Promising Types of Stem Cells for Therapy are Embryonic Stem Cells

Page 87: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

The Inner Cell Mass is the Source of Embryonic Stem Cells

The embryo is destroyed by separating it into individual cells for the collection of ICM cells.

Page 88: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Some Thorny Ethical Questions

Is it ethical to harvest embryonic stem cells from the “extra” embryos created during in vitro fertilization?

Are these masses of cells a human?

Page 89: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

The Promise and Possible Perils of Stem Cells

Page 90: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Additional Potential Dilemmas – Therapeutic Cloning to Obtain Matched Embryonic Stem Cells

Cells from any source other than you or an identical twin present the problem of rejection.

If so, how can matched embryonic stem cells be obtained?

A cloned embryo of a person can be made, and embryonic stem cells harvested from these clones.

Cultured mouse embryonic stem cells.

Page 91: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Therapeutic Cloning

Is there any ethical difference between therapeutic and reproductive cloning?

Page 92: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

HIV/Pregnancy Tests

Done using body’s own immune response ELISA test

Cells naturally have antigens on their outer surface.

The antibodies respond to the antigens in an immune response

Antibodies and antigens fit together “like a glove”

Page 93: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy
Page 94: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy
Page 95: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

http://www.elispot-analyzers.de/english/elisa-animation.html

Animation

Page 96: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

http://www.genome.gov/Pages/EducationKit/download.htmlDNA technology video

Page 97: DNA Technologies. Biotechnology Dolly and surrogate Mom Genetically modified rice. Embryonic stem cells and gene therapy

Online Tools

• Buying primers (custom oligos): http://www.qiagen.com/• 2x2 sequence comparisons: http://www2.igh.cnrs.fr/bin/align-

guess.cgi• n x n sequence comparisons:

http://www.genebee.msu.su/services/malign_reduced.html• Sequence manipulation:

http://arbl.cvmbs.colostate.edu/molkit/manip/index.html• Sequence translation:

http://arbl.cvmbs.colostate.edu/molkit/translate/index.html• Restriction Enzymes: http://rebase.neb.com/rebase/rebase.html• Restriction sites:

http://www.ccsi.com/firstmarket/cutter/cut2.html• Sequence searches: http://www.ncbi.nlm.nih.gov/blast/• Other tools: http://molbiol-tools.ca/