Page 9 Page 10 Gel electrophoresis is a
- Slides: 43
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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 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
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 3. QTL mapping 4. Maker-assisted selection for breeding program Page 15
Restriction Enzymes Page 16
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PCR-based methods
Random Amplified polymorphic DNA (RAPDs) Page 20
Aplifed Fragment length Polymorphism (AFLPs) Page 21
Gel for AFLP markers Page 22
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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 : (CTG)4 ACTCACTC : (ACTC)4
Microsatellites
Applications of Molecular Markers • Genetic diversity and fingerprinting • Inheritance and mapping – Simply inherited traits – Quantitative traits (dissection into QTL)
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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 populations consisting of thousands of plants MARKER-ASSISTED SELECTION (MAS) Method whereby phenotypic selection is based on DNA markers
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Marker-based gene identification and isolation
Particle Gun Page 37
Thank you mahmoud_zabalawy@yahoo. com Page 42
General Discussion Page 43
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