Page 9 Page 10 Gel electrophoresis is a

  • Slides: 43
Download presentation

Page 9

Page 9

Page 10

Page 10

Gel electrophoresis • is a technique used for the separation of DNA, RNA or

Gel electrophoresis • is a technique used for the separation of DNA, RNA or protein molecules using an electric current applied to a gel matrix. • It is usually performed for analytical purposes, such as RFLP , PCR, Page 11

 ﺃﻬﻢ ﻃﺮﻕ ﺍﻟﻤﻌﻠﻤﺎﺕ ﺍﻟﻮﺭﺍﺛﻴﺔ Restriction Fragment Length Polymorphisms (RFLP) Minisatellites or Variable Number

ﺃﻬﻢ ﻃﺮﻕ ﺍﻟﻤﻌﻠﻤﺎﺕ ﺍﻟﻮﺭﺍﺛﻴﺔ Restriction Fragment Length Polymorphisms (RFLP) Minisatellites or Variable Number of Tandem Repeats (VNTR) Internal Transcribed Spacer regions of nuclear ribosomal genes (ITS) Sequence-Tagged Sites (STS) Microsatellites, Simple Sequence Repeat (SSR), Amplified Sequence Length Polymorphism (ASLP) Sequence Characterized Amplified Region (SCAR) Cleaved Amplified Polymorphic Sequence (CAPS) Single-Strand Conformation Polymorphism (SSCP) Denaturing Gradient Gel Electrophoresis (DGGE) Thermal Gradient Gel Electrophoresis (TGGE) Heteroduplex Analysis (HDA) Denaturing High Performance Liquid Chromatography (DHPLC) Multiple Arbitrary Amplicon Profiling (MAAP) Random Amplified Polymorphic DNA (RAPD) DNA Amplification Fingerprinting (DAF) Arbitrarily Primed Polymerase Chain Reaction (AP-PCR) Inter-Simple Sequence Repeat (ISSR) Single Primer Amplification Reaction (SPAR) Directed Amplification of Minisatellite DNA (DAMD) Amplified Fragment Length Polymorphism (AFLP) Selectively Amplified Microsatellite Polymorphic Loci (SAMPL) Page 12

Advances in molecular markers Page 13

Advances in molecular markers Page 13

Examples of Techniques to detect DNA polymorphism (DNA markers) i) Non-PCR based marker -

Examples of Techniques to detect DNA polymorphism (DNA markers) i) Non-PCR based marker - RFLP ii) PCR-based markers - RAPD - AFLP - SSR Page 14

Restriction Fragment Length Polymorphism (RFLP) Applications of RFLP 1. DNA Fingerprinting 2. Genetic mapping

Restriction Fragment Length Polymorphism (RFLP) Applications of RFLP 1. DNA Fingerprinting 2. Genetic mapping 3. QTL mapping 4. Maker-assisted selection for breeding program Page 15

Restriction Enzymes Page 16

Restriction Enzymes Page 16

Page 17

Page 17

Page 18

Page 18

PCR-based methods

PCR-based methods

Random Amplified polymorphic DNA (RAPDs) Page 20

Random Amplified polymorphic DNA (RAPDs) Page 20

Aplifed Fragment length Polymorphism (AFLPs) Page 21

Aplifed Fragment length Polymorphism (AFLPs) Page 21

Gel for AFLP markers Page 22

Gel for AFLP markers Page 22

Page 23

Page 23

Page 24

Page 24

Genetic diversity among wheat genotypes by means AFLP Markers Page 25

Genetic diversity among wheat genotypes by means AFLP Markers Page 25

Microsatellites or Simple Sequence Repeats (SSRs) Description: AAAAAA : (A)11 GTGTGT : (GT)6 CTGCTG

Microsatellites or Simple Sequence Repeats (SSRs) Description: AAAAAA : (A)11 GTGTGT : (GT)6 CTGCTG : (CTG)4 ACTCACTC : (ACTC)4

Microsatellites

Microsatellites

Applications of Molecular Markers • Genetic diversity and fingerprinting • Inheritance and mapping –

Applications of Molecular Markers • Genetic diversity and fingerprinting • Inheritance and mapping – Simply inherited traits – Quantitative traits (dissection into QTL)

Page 29

Page 29

Page 30

Page 30

Page 31

Page 31

Page 32

Page 32

Applications of Molecular Markers • As selection tools: Marker -Assisted Selection

Applications of Molecular Markers • As selection tools: Marker -Assisted Selection

MARKER-ASSISTED BREEDING Susceptible P 1 x P 2 Resistant F 1 F 2 large

MARKER-ASSISTED BREEDING Susceptible P 1 x P 2 Resistant F 1 F 2 large populations consisting of thousands of plants MARKER-ASSISTED SELECTION (MAS) Method whereby phenotypic selection is based on DNA markers

Page 35

Page 35

Marker-based gene identification and isolation

Marker-based gene identification and isolation

Particle Gun Page 37

Particle Gun Page 37

Thank you mahmoud_zabalawy@yahoo. com Page 42

Thank you mahmoud_zabalawy@yahoo. com Page 42

General Discussion Page 43

General Discussion Page 43