Applications of DNA technology

Slides:



Advertisements
Similar presentations
13-2 Manipulating DNA.
Advertisements

Recombinant DNA Technology for the non- science major.
DNA Technology- Cloning, Libraries, and PCR 17 November, 2003 Text Chapter 20.
6.3 Advanced Molecular Biological Techniques 1. Polymerase chain reaction (PCR) 2. Restriction fragment length polymorphism (RFLP) 3. DNA sequencing.
Manipulating DNA Genetic Engineering uses the understanding of the properties of DNA to study and change DNA sequences in living organisms – Invitro… in.
Chapter 20~DNA Technology & Genomics. Who am I? Recombinant DNA n Def: DNA in which genes from 2 different sources are linked n Genetic engineering:
Chapter 19 – Molecular Genetic Analysis and Biotechnology
1 Genetics Faculty of Agriculture Instructor: Dr. Jihad Abdallah Topic 13:Recombinant DNA Technology.
DNA Technology Chapter 20.
Slide 1 of 32 Copyright Pearson Prentice Hall Biology.
Technological Solutions. In 1977 Sanger et al. were able to work out the complete nucleotide sequence in a virus – (Phage 0X174) This breakthrough allowed.
Manipulating DNA.
Module 1 Section 1.3 DNA Technology
13-1 Changing the Living World
Background Genetic disorders are often the result of gene mutations. People with a mutant allele often have a family history of the disease. It is important.
Genetics 6: Techniques for Producing and Analyzing DNA.
Review from last week. The Making of a Plasmid Plasmid: - a small circular piece of extra-chromosomal bacterial DNA, able to replicate - bacteria exchange.
Highlights of DNA Technology. Cloning technology has many applications: Many copies of the gene are made Protein products can be produced.
GENETIC ENGINEERING CHAPTER 20
Bioinformatics & Biotechnology Lecture 1 Sequencing BLAST PCR Gel Electrophoresis.
Human Genomics. Writing in RED indicates the SQA outcomes. Writing in BLACK explains these outcomes in depth.
6.3 Advanced Molecular Biological Techniques 1. Polymerase chain reaction (PCR) 2. Restriction fragment length polymorphism (RFLP) 3. DNA sequencing.
Chapter 10: Genetic Engineering- A Revolution in Molecular Biology.
Locating and sequencing genes
Manipulating DNA. Scientists use their knowledge of the structure of DNA and its chemical properties to study and change DNA molecules Different techniques.
Molecular Basis for Relationship between Genotype and Phenotype DNA RNA protein genotype function organism phenotype DNA sequence amino acid sequence transcription.
Advantages of STR Analysis
{ Genetics Review.  Involves several different genes for one trait like eye color, skin tone and color, height (humans), wheat kernel color  These are.
BIOTECHNOLOGY DNA is now being easily manipulated. Molecular biologists analyze and alter genes and their respective proteins. Recombinant DNA is DNA from.
Molecular Genetic Technologies Gel Electrophoresis PCR Restriction & ligation Enzymes Recombinant plasmids and transformation DNA microarrays DNA profiling.
DNA Technology Ch. 20. The Human Genome The human genome has over 3 billion base pairs 97% does not code for proteins Called “Junk DNA” or “Noncoding.
MOLECULAR GENETICS TECHNIQUES. Molecular Genetics Technologies i.Polymerase chain reaction ii.DNA/Genomic sequencing iii.Gel electrophoresis iv.Restriction.
Chapter 20 DNA Technology and Genomics. Biotechnology is the manipulation of organisms or their components to make useful products. Recombinant DNA is.
FOOTHILL HIGH SCHOOL SCIENCE DEPARTMENT Chapter 13 Genetic Engineering Section 13-2 Manipulating DNA.
Gene Technologies and Human ApplicationsSection 3 Section 3: Gene Technologies in Detail Preview Bellringer Key Ideas Basic Tools for Genetic Manipulation.
Chapter 14 GENETIC TECHNOLOGY. A. Manipulation and Modification of DNA 1. Restriction Enzymes Recognize specific sequences of DNA (usually palindromes)
The genetic engineers toolkit A brief overview of some of the techniques commonly used.
Title: Studying whole genomes Homework: learning package 14 for Thursday 21 June 2016.
Ch 15 DNA Technology/ Genetic Engineering
Copyright Pearson Prentice Hall
Chapter 7 Recombinant DNA Technology and Genomics
DNA Technologies (Introduction)
Genomics Sequencing genomes.
Recombinant DNA Technology
copying & sequencing DNA
Section 3: Gene Technologies in Detail
Genetic Research and Biotechnology
AMPLIFYING AND ANALYZING DNA.
DNA Technology.
AMPLIFYING AND ANALYZING DNA.
Chapter 20 – DNA Technology and Genomics
Relationship between Genotype and Phenotype
How are areas of DNA that don’t code for proteins (genes) used by our cells? How can we make use of these areas?
DNA Technology.
The student is expected to: (6H) describe how techniques such as DNA fingerprinting, genetic modifications, and chromosomal analysis are used to study.
Copyright Pearson Prentice Hall
Sequencing and Copying DNA
Copyright Pearson Prentice Hall
Recombinant DNA Unit 12 Lesson 2.
DNA and the Genome Key Area 8a Genomic Sequencing.
Copyright Pearson Prentice Hall
Copyright Pearson Prentice Hall
Copyright Pearson Prentice Hall
Polymerase Chain Reaction PCR
Relationship between Genotype and Phenotype
Copyright Pearson Prentice Hall
SBI4U0 Biotechnology.
Forensic DNA Sadeq Kaabi
Using the DNA Sequence Knowing the sequence of an organism’s DNA allows researchers to study specific genes, to compare them with the genes of other organisms,
Polymerase Chain Reaction (PCR) & DNA SEQUENCING
Presentation transcript:

Applications of DNA technology Human Genome Project Human Therapeutics Forensic Uses Agriculture

History Late 1980’s idea was proposed Predicted it would take 15 years Cost about $200 million per year $1 per base pair Officially began in 1990 26 June 2000 joint announcement from Blair and Clinton ‘the draft complete’ Joint publication in Nature and Science 12 Feb 2001 14 Apr 2003 – The finished human genome Discuss here the links between the public and private sector. Worth noting that the Wellcome Trust through the creation of the Sanger Institute was responsible for sequencing 1/3 of the DNA sequence.

Why? ‘If I were to study the carburettor of my car engine, even in exquisite detail, I would still have no idea about the overall function of the engine, much less the entire car. To understand what an engine is for, and how it works, I’d need to study the whole thing – I’d need to place the carburettor in context, as one function part among many. The same is true of genes…..’ James Watson

Method Genetic Mapping Physical Mapping DNA sequencing Identifies relative positions of genes E.g. Gene 2 lies between genes 1 and 3 Physical Mapping Absolute positions of genes on chromosomes E.g. Gene 2 is 1 million bp from gene 1 DNA sequencing Actual ATCG combinations Mention the understanding that computers and technology would improve over time therefore speed would increase and cost decrease. Also mention the practise gained by sequencing genomes of smaller organisms such as the nematode, yeast and mice. Compare naming all the houses between edinburgh and London in the correct order. Genetic mapping looks at the towns and gets them in the correct order, physical mapping knows exactly how far apart these towns are from one another, then get down to looking at the houses and where they lie in relation to the sequence.

Polymerase Chain Reaction Critical to the HGP was the ability to create large amounts of DNA for sequencing. PCR is a process to amplify DNA DNA is heated to 95oC The DNA is denatured causing the two strands to separate A primer (short length of DNA) binds (or anneals) to the template strands [after the solution is cooled] Complementary DNA strands form [through the action of DNA polymerase] Lead into this by explaining that we will look at the methods in more detail, but in the end it wasn’t the invention of new methods that sped up the sequencing process, but the automation of old processes. Amplification of DNA used to be done by inserting the required DNA sequence into plasmids of bacteria and then culturing the bacteria. PCR was a process invented by Kary Mullis. Early problems with the method included the need to add more polymerase after each cycle, as DNA polymerase is denatured at 95oC. This problem was solved by thermus aquaticus bacteria (taq polymerase)

Discuss the need for primers (DNA polymerase can only act on double stranded molecules). Highlight how primers are sequences created artificially. We can sequence only the gene wanted from a larger chunk of DNA just by choosing the primer correctly

Genetic Mapping Genetic mapping utilises cross over frequencies between known genetic markers. A genetic marker is any sequence of genome that shows difference between individuals. It could be A gene A Microsatellite or Short Tandem Repeat Differences in genes didn’t give enough information – often they were looking for mutations in the gene and therefore looked for families that were carriers and sufferers of genetic diseases. Turned to STRs which are frequent throughout the genome to give markers.

Microsatellites (STRs) Repeating sequences 2 – 4 nucleotide bases found in mostly in introns (‘junk’ DNA) The number of repeats varies from person to person, but follow patterns of Mendalian inheritance These sequences can be identified using probes. A probe is A short, single stranded sequence of DNA Complementary to DNA base sequence ‘Tagged’ – radioactively or using fluorescent dye Radioactive tags show up with exposure to photographic film. Fluorescent dye shows up when exposed to UV light.

Recombinance / Cross Over Frequency What do you remember about linked genes from Higher. What information do recombinant individuals give us about the relative position of genes? Cross over frequency is the number of recombinant individuals / total number of offspring. Recombinants are different from parental types.

Physical Mapping Use of restriction enzymes Restriction enzymes belong to the group ‘endonucleases’ or nucleases. They cut DNA at specific sequences. Chromosomes can therefore be cut into sequences of different lengths. By using combinations of restriction enzymes and working out the size of the fragments, a pattern of recognition sites in the DNA can be pieced together. Note that there are many different types of physical maps at different resolutions – the ultimate being a total DNA sequence. Recall nucleases break phosphodiester bonds (i.e. break down nucleic acids). Endonucleases are isolated from bacteria, in which it is believed they are used in cellular defences against invading viruses.

Gel electrophoresis Uses physical properties of DNA (size and charge) to separate molecules Gel electrophoresis involves running an electric current through an agarose gel. DNA is loaded into wells at the negative end. DNA is repelled, and moves through the agarose gel at different speeds depending on the size of a fragment. A Molecular weight marker (of known fragment size) is also used. Samples can be compared to this marker to work out their size. Note the many different use of gel electrophoresis, but highlight its importance in this case in determining the size of the fragments.

Physical Mapping cont… M = molecular weight marker 1 = undigested sample 2 = digested with NotI 3 = digested with BamHI 4 = digested with NotI and BamHI 5 = something totally different

Physical Mapping cont…

Physical Mapping cont….

DNA Sequencing Sanger method (aka dideoxy chain-termination method) An unknown DNA template strand is replicated using a primer (to initiate replication), DNA polymerase, nucleotide bases and one of 4 dideoxy nucleotide bases (ddA, ddG, ddC and ddT)

DNA sequencing cont… If a ddNTP is inserted instead of a normal nucleotide DNA replication stops at that point (chain termination) When this experiment is repeated with other dNTPs DNA molecules differing in length by 1 bp are created.

DNA sequencing cont.. These are then run on an electrophoresis gel Note: The smallest pieces at the start of the sequence are at the bottom. Therefore the sequence is read bottom to top Note that these are usually tagged with a radioactive sample. These can be compared.

DNA sequencing cont… Talk about newly automated methods – these involve tagging with a fluoresent dye. A computer gives a print out of the different bases.

Comparing Genomes Other genomes have been sequenced in conjunction with the human genome, and have continued. There are a large number of similarities between genomes. Genes found in a wide range of organisms are known as homologous genes. These are used to study function of genes.

Learning Activities Look at the arrangements document to clarify what information is required. Read DART pg 73 – 81. Read the Monograph pg 67 – 79 Scholar – 8 Internet research Worksheets