Restriction Enzymes and Gel Electrophoresis. What is a Palindrome? A palindrome is anything that reads the same forwards and backwards: A palindrome is.

Slides:

Advertisements

Similar presentations

James Chappell & Cheuk Ka Tong

Advertisements

UNIT 2 MANIPULATION OF DNA AND GENE ISOLATION LECTURES: 9. DNA Cloning and Library Construction 10. Isolating Genes.

RESTRICTION ENZYMES & GEL ELECTROPHORESIS ANALYSIS OF PRECUT LAMBDA DNA.

Introduction to Techniques

Basic methods in genetics PCR; Polymerase Chain Reaction Restriction enzyme digestions Gel electrophoresis.

Restriction Enzymes. Molecular Scissors Restriction enzymes are like molecular scissors. They scan the DNA strand looking for particular sequences of.

6.1 Biotechnological Tools and Techniques Recombinant DNA & Gel electrophoresis.

V) BIOTECHNOLOGY.

Bacterial Transformation

Restriction Enzymes DNA is a long chain of nucleotides. AluI recognises AGCT, and fixes itself on its target sequence. It cuts after the G to give Blunt.

Restriction Enzymes and Gel Electrophoresis

Introduction to Bioinformatics Molecular Biology Tools.

Restriction Enzymes.

& Gel Plasmid Electrophoresis Mapping.

©2000 Timothy G. Standish Restriction Enzyme Digestion Timothy G. Standish, Ph. D.

Part 1. Gel electrophoresis DNA is negatively charged (because of phosphate backbone) DNA will be attracted to positively charged poles and repelled from.

Restriction Digestion and Analysis of Lambda DNA Kit

Genetic Engineering DNA technology. Palindromes  mom  wow  race car  straw warts  never odd or even  stella won no wallets  was it a car or a cat.

(RFLP Electrophoresis)

Quickie Intro to DNA Technologies

Happy Monday! Checking off: Notes on Ch 20.1, 20.2 With your group, discuss what you know about these: – Methylation and Acetylation – Genetic Engineering/Biotechnology.

Basic methods in genetics PCR; Polymerase Chain Reaction Restriction enzyme digestions Gel electrophoresis.

Restriction Enzymes. Restriction Endonucleases Also called restriction enzymes “molecular scissors” discovered in in bacteria Restriction enzymes is an.

KEY CONCEPT Biotechnology relies on cutting DNA at specific places.

Molecular Genetics Lab Review. Bacterial Transformation Genetic transformation—host organism takes in and expresses foreign DNA Genetic engineering—manipulation.

Part One BIOTECHNOLOGICAL TOOLS & TECHNIQUES. What is biotechnology? Applied biology genetics; molecular biology; microbiology; biochemistry Uses living.

Chapter 9: Genetic Engineering

(RFLP Electrophoresis)

Chapter 20 DNA Technology & Genomics. Genetic engineering Manipulation of genetic material for practical purposes has begun industrial revolution in biotechnology.

DNA fingerprinting is not taking someone’s fingerprint. It is cutting up a DNA strand and separating them by size.

Chapter 4 Molecular Cloning Methods Jay D. Hunt, Ph.D. Department of Biochemistry and Molecular Biology CSRB 4D

DNA Fingerprinting. Introduction to DNA Fingerprinting Technicians in forensic labs are often asked to do DNA profiling or “fingerprinting” Restriction.

Restriction Enzyme Digest October 8, Restriction Enzymes.

Restriction Enzymes Biotechnology Fall 2013.

Gel Electrophoresis By Erin Martin & Satya Moolani.

nome/program.html.

The genetic engineers toolkit A brief overview of some of the techniques commonly used.

RESTRICTION ENZYMES & GEL ELECTROPHORESIS FORENSIC DNA FINGERPRINTING.

Biotechnology I. POINT > Define what restriction enzymes are POINT > Describe how restriction enzymes cut DNA POINT > Show how restriction enzymes facilitate.

1 DNA Technology Part I: Restriction Enzymes & Electrophoresis Created by: C. Massengale Edited by: Beth Walker.

Palindrome, Restriction Enzyme, Sticky Ends GAATTC G AATTC G Sticky Ends (Cohesive Ends) EcoRI CIVIC, Madam Get An Apple To The Class G AATTCG Arber, Nathans,

Electrophoresis is a molecular technique that separates nucleic acids and proteins based on Size and +-+ Charge +-+

TOOLS OF GENETIC ENGINEERING. There are a number of tools used in genetic engineering. One of them are enzymes.

Restriction Enzymes. Discovery  In 1962, Werner Arber, a Swiss biochemist, provided the first evidence for the existence of "molecular scissors" that.

Restriction Enzymes. Molecular Scissors Restriction enzymes are molecular scissors.

Forensic Investigation

Biotechnology Restriction Enzymes 4/16/2018.

Restriction Enzymes and Digestion

Bacterial Transformation

Chapter 4 Recombinant DNA Technology

BIOTECHNOLOGICAL TOOLS & TECHNIQUES

Thanks to Cal State LA! Kuhn/Pickett

Biotechnology Notes Chapter 9.

KEY CONCEPT Biotechnology relies on cutting DNA at specific places.

Restriction Enzymes and Digestion

Restriction Enzymes & Electrophoresis

Forensic Investigation

Recombinant DNA Unit 12 Lesson 2.

Restriction Enzymes.

Restriction Digestion and Analysis of Lambda DNA Kit

KEY CONCEPT Biotechnology relies on cutting DNA at specific places.

Simulating Genetic Screening

Genetics and Biotechnology

KEY CONCEPT Biotechnology relies on cutting DNA at specific places.

Biotechnology: Restriction Enzyme Analysis of DNA

KEY CONCEPT Biotechnology relies on cutting DNA at specific places.

Restriction Digestion and Analysis of Lambda DNA Kit

KEY CONCEPT Biotechnology relies on cutting DNA at specific places.

KEY CONCEPT Biotechnology relies on cutting DNA at specific places.

Presentation transcript:

Restriction Enzymes and Gel Electrophoresis

What is a Palindrome? A palindrome is anything that reads the same forwards and backwards: A palindrome is anything that reads the same forwards and backwards: English palindromes: English palindromes: mom mom dad dad Rats live on no evil star. Rats live on no evil star.

DNA Palindromes Palindromes are defined as any double stranded DNA which when read 5’ to 3’ and 3’ to 5’, they read the same. Palindromes are defined as any double stranded DNA which when read 5’ to 3’ and 3’ to 5’, they read the same. The EcoRI cutting site: 5'-GAATTC-3' 3'-CTTAAG-5' The HindIII cutting site: 5'-AAGCTT-3' 3'-TTCGAA-5'

Restriction Enzyme Review DNA scissors or cuts DNADNA scissors or cuts DNA Two types of cuts:Two types of cuts: Blunt endsBlunt ends Sticky endsSticky ends Each enzyme has a specific palindromic sequence that it recognizesEach enzyme has a specific palindromic sequence that it recognizes

How many Fragments? DNA: AGTCGTAGCTCGATC CC GGGCTGATCGAA TCAGCATCGAGCTAG GG CCCGACTAGCTT Restriction Enzyme: Sma I cuts at …C C C G G G… …G GG C C C… How many fragments? AGTCGTAGCTCGATCCC GGG CTGATCGAA TCAGCATCGAGCTAGGG CCC GACTAGCTT Restriction site Cut site

Step 2: Gel Electrophoresis Separates DNA (or RNA or Protein) fragments based on size and chargeSeparates DNA (or RNA or Protein) fragments based on size and charge Each sample goes into its own well and then has its own laneEach sample goes into its own well and then has its own lane Larger fragments move slower through the gelLarger fragments move slower through the gel

Electrophoresis = movement of charged particles using an electric fieldElectrophoresis = movement of charged particles using an electric field DNA is negatively charged so it moves towards the positive end of the boxDNA is negatively charged so it moves towards the positive end of the box Step 2: Gel Electrophoresis

Length of Fragments Number of base pairsNumber of base pairs 1 Base Pair = 2 Nucleotides1 Base Pair = 2 Nucleotides Examples: How many base pairs?Examples: How many base pairs? DNA Ladder = sample made up of DNA with known lengths Blunt ends: ATGTC TACAG Sticky ends: AGCTTGTA ACATTCGA ATAT

Gel Electrophoresis Wells

Gel Electrophoresis Wells

Gel Electrophoresis + - Direction of DNA Travel Wells Small Large DNA Ladder

G CTTAA AATTC G 1 Digestion 2 Annealing of sticky ends 3 Ligation Ligase G CTTAA AATTC G EcoRI R. E.s and DNA Ligase Can be used to make recombinant DNA GAATTC CTTAAG GAATTC CTTAAG G CTTAA AATTC G 4 Recombinant DNA