Microsatelit
Download as PPTX, PDF4 likes794 views
This document discusses microsatellites and molecular markers. It begins by defining microsatellites as simple sequence repeats between 1-6 base pairs long. It then discusses different types of markers including morphological, biochemical, chromosomal, and genetic markers. The main part of the document focuses on different molecular markers used in genetics including RFLP, RAPD, AFLP, microsatellites, DNA fingerprinting, and SNPs. It provides details on what each marker is and how it is detected. The document concludes by discussing applications of molecular markers in areas like gene mapping, disease diagnosis, evolution studies, and animal breeding/selection.
Microsatelit
- 1. Microsatelit Oleh: Awari Susanti BP: 1320422015 Biologi Molekuler PROGRAM PASCASARJANA BIOLOGI FAKULTAS MATEMATIKA DAN ILMU PENGETAHUAN ALAM UNIVERSITAS ANDALAS PADANG,2014
- 2. What is microsatellite • Simple Sequence Repeats (SSR) • 1-6 bp long
- 3. What is Marker? Morphological Biochemical Chromosomal Genetic Marker is a piece of DNA molecule that is associated with a certain trait of a organism Types of Markers
- 4. Animals are selected based on appearance Eg. PIGMENTATION Disadvantage: lack of polymorphism
- 5. Animals are selected based on biochemical properties Eg. Hb, AMYLASE, BLOOD GROUPS ETC. Disadvantage: Sex limited Age dependent Influenced by environment It covers less than 10% of genome
- 6. Animals are selected based on structural & numerical variations Eg. Structural and Numerical Variations Structural- Deletions, Insertions etc. Numerical- Trisomy, Monosomy, Nullysomy Disadvantage: low polymorphism
- 7. Molecular Marker • Revealing variation at a DNA level • Characteristics: – Co-dominant expression – Nondestructive assay – Complete penetrance – Early onset of phenotypic expression – High polymorphism – Random distribution throughout the genome – Assay can be automated
- 9. DNA isolated from any tissue eg. Blood, hair etc. DNA isolated at any stage even during foetal life DNA has longer shelf-life readily exchangeable b/w labs Analysis of DNA carried out at early age/ even at the embryonic Stage irrespective of sex.
- 10. Molecular Markers Single locus marker Multi-locus marker RFLP Microsatellite STS DNA Fingerprinting AFLP RAPD
- 11. M 1 2 3 4 5 6 7 8 9 10 11 –ve M Single locus marker
- 13. Randomly Amplified Polymorphic DNA (RAPD) • PCR based marker with 10-12 base pairs • Random amplification of several fragments • Amplified fragments run in agarose gel detected by EtBr • Unstable amplification leads to poor repeatability
- 15. Restriction Fragment Length Polymorphism (RFLP) • Genomic DNA digested with Restriction Enzymes • DNA fragments separated via electrophoresis and transfer to nylon membrane • Membranes exposed to probes labelled with P32 via southern hybridization • Film exposed to X-Ray
- 17. Amplified Fragment Length Polymorphism (AFLP) • Restriction endonuclease digestion of DNA • Ligation of adaptors • Amplification of ligated fragments • Separation of the amplified fragments via electrophoresis and visualization • AFLPs have stable amplification and good repeatability
- 19. SSR: Simple Sequence Repeat or Microsatellite • PCR based markers with 18-25 base pair primers • SSR polymorphisms are based on no. of repeat units and are hypervariable • SSRs have stable amplification and good repeatability • SSR are easy to run and automate
- 21. DFP: DNA finger printing • DNA extraction from individual • Amplification of markers • Electrophoresis separation of markers • Visualization of markers • Scoring of markers for each individual • Data analysis
- 23. Features RFLP PCR-RFLP DFP RAPD Microsatellite SNP Detection method Hybridization PCR Hybridization PCR PCR PCR Type of probe/primer used g DNA/ cDNA sequence of structural genes Sequence specific primers Mini satellite synthetic oligos Arbitrarily design primer Sequence specific primers Sequence specific primers Requirement of radioactivity Yes No/Yes Yes No/Yes No/Yes No/Yes Extant of genomic coverage Limited Limited Extensive Extensive Extensive Extensive Degree of polymorphisms Low Low High Medium to High High High Phenotype expression Co dominant Co dominant Co dominant Co dominant/ Dominant Dominant Co dominant Possibility of No Yes No Yes Yes Yes
- 24. • Gene mapping • Pre and post natal diagnosis of diseases • Anthropological and molecular evolution studies
- 25. Short Range Application • Parentage determination • Genetic distance estimation • Determination of twin zygosity & freemartins • Sexing of pre-implanted embryos • Identification of disease carries
- 26. Long Range Applications • Gene mapping & mapping of QTL by linkage • Marker assisted selection