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Thematic presentation:
MOLECULE TECHNIQUES
MOLECULE TECHNIQUES
Members
1. Hoang Thi Huyen Trang 11126237 DH11SH 2. Nguyen Thi Hong Phuc 3. Le Dang Huynh Tram 4. Le Nguyen Thao Ly 5. Le Nhat Tan 11126183 DH11SH 11126241 DH11SH 11126308 DH11SH 11126321 DH11SH
Sections
Hybridization PCR
Electrophoresis
Molecule techniques
Principle
The nucleic acid hybridization is the process where in two DNA or RNA single chains from different biological sources, make the double catenar configuration, based on nucleotide complementarity and of contingen sequence homology of the two sources, resulting DNA-DNA, RNA-RNA or DNARNA hybrids.
Principle
In most cases, the purpose of the hybridization techniques is identification or localization of certain nucleic acid sequences in the genome of some species. Two basic notions are used: the target molecule representing the DNA, RNA or protein sequence that should be identified and the probe molecule who identify the target, by hybridization.
Categories
Hybridization stages
1. Probe synthesis 2. Probe marking
6. Molecular hybridization
Southern Blot
Stages
Modifications
First nylon membranes are less fragile than nitrocellulose sheets
Certain conditions the transferred DNA becomes covalently bound to the membrane during the transfer process
Nylon membranes efficiently bind DNA fragments down to 50 bp in length, where as nitrocellulose membranes are effective only with molecules longer than 500 bp
RFLP
In molecular biology, restriction fragment length polymorphism is a technique that exploits variations in homologous DNA sequences. It refers to a difference between samples of homologous DNA molecules that come from differing locations of restriction enzyme sites, and to a related laboratory technique by which these segments can be illustrated. In RFLP analysis, the DNA sample is broken into pieces (digested) by restriction enzymes and the resulting restriction fragments are separated according to their lengths by gel electrophoresis.
Applications
Analysis of RFLP variation in genomes was a vital tool in genome mapping and genetic disease analysis
RFLP analysis was also the basis for early methods of Genetic fingerprinting
Microarray
A microarray is a multiplex lab-on-a-chip. It is a 2D array on a solid substrate (usually a glass slide or silicon thin-film cell) that assays large amounts of biological material using high-throughput screening methods. The concept and methodology of microarrays was first introduced and illustrated in antibody microarrays (also referred to as antibody matrix) in 1983 in a scientific publication and a series of patents
Applications
Help researchers to learn more about many different diseases including heart disease, mental illness and infectious diseases
Additionally, by examining the differences in gene activity between untreated and treated tumor cells.
The polymerase chain reaction (PCR) is a biochemical technology in molecular biology to amplify a single or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence.
The method relies on thermal cycling, consist of cycles of repeated heating and cooling of the reaction for DNA melting and enzymatic replication of the DNA
Components
DNA template that contains the DNA region (target) to be amplified. Two primers that are complementary to the 3' (three prime) ends of each of the sense and anti-sense strand of the DNA target. Taq polymerase or another DNA polymerase with a temperature optimum at around 70 C.
Components
Deoxynucleoside triphosphates (dNTPs, sometimes called "deoxynucleotide triphosphates)
Buffer solution, providing a suitable chemical environment for optimum activity and stability of the DNA polymerase.
Divalent cations, magnesium or manganese ions (Mg2+, Mn2+)
PCR stages
Application of PCR
Figure : Electrophoresis of PCR-amplified DNA fragments. (1) Father. (2) Child. (3) Mother. The child has inherited some, but not all of the fingerprint of each of its parents, giving it a new, unique fingerprint.
Application of PCR
Electrophoresis
Electrophoresis is the motion of dispersed particles relative to a fluid under the influence of a spatially uniform electric field. This electrokinetic phenomenon was observed for the first time in 1807 by Ferdinand Frederic Reuss, who noticed that the application of a constant electric field caused clay particles dispersed in water to migrate. It is ultimately caused by the presence of a charged interface between the particle surface and the surrounding fluid. It is the basis for a number of analytical techniques used in biochemistry for separating molecules by size, charge, or binding affinity.
Applications
1 DNA Analysis
Protein Analysis
Antibiotics Analysis
Vaccine Analysis
References
http://www.hc-sc.gc.ca/sr-sr/biotech/aboutapropos/molecul_tech-eng.php
http://en.wikipedia.org/wiki/Polymerase_chai n_reaction
http://en.wikipedia.org/wiki/Electrophoresis
http://en.wikipedia.org/wiki/Hybridisation