PCR --Polymerase Chain Reaction
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Polymerase Chain Reaction, PCR, DNA Amplification, Thermocycler, Taq Polymerase, Polymerase Chain Reaction, PCR, DNA Amplification, Thermocycler, Taq Polymerase, Polymerase Chain Reaction, PCR, DNA Amplification, Thermocycler, Taq Polymerase, Polymerase Chain Reaction, PCR, DNA Amplification, Thermocycler, Taq Polymerase
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Types of PCR
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Nucleic acid amplification techniques involve synthesizing many copies of DNA or RNA from a template. They are classified into target amplification, probe amplification, and signal amplification. Polymerase chain reaction (PCR) is a commonly used target amplification technique that exponentially amplifies target DNA sequences in vitro using DNA polymerase. PCR has many applications including DNA sequencing, bacterial cloning, and detecting pathogens. It involves thermal cycling between denaturation, annealing of primers, and extension steps. Variations of PCR like real-time PCR, nested PCR, and digital PCR have further expanded its uses.Polymerase chain reaction
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The document discusses the Polymerase Chain Reaction (PCR) technique. It was invented by Kary Mullis in 1985. PCR allows for targeted amplification of specific DNA sequences. It works by using DNA polymerase to make copies of a targeted region of DNA defined by primer sequences. Through repeated heating and cooling cycles, millions of copies of the target DNA can be produced, allowing it to be analyzed. Taq polymerase, an enzyme from thermophilic bacteria, is used as it is heat-stable and allows the reaction to take place.PCR PPT
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PCR (polymerase chain reaction) was invented in 1987 by Kary Mullis to specifically amplify a single DNA sequence. It takes advantage of DNA replication by using DNA polymerase, primers, and thermocycling to exponentially amplify the target sequence. PCR is now widely used in applications such as cloning, forensics, disease detection, and more due to its ability to amplify small amounts of DNA.Polymerase chain reaction Pranav
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Polymerase chain reaction is a technique used in molecular biology to amplify a single copy or a few copies of a segment of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequenceQ pcr
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The document discusses Polymerase Chain Reaction (PCR), including its history, components, process, applications, and improvements. It was invented in 1983 by Kary Mullis, who received the Nobel Prize for it. PCR exponentially amplifies a specific DNA sequence using DNA polymerase, primers, and repeated heating and cooling cycles. It has numerous applications including DNA sequencing, pathogen detection, genetic analysis, and more. Later developments improved PCR, such as nested PCR, quantitative PCR, and sequencing technologies.Telomere
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The polymerase chain reaction (PCR) is an in vitro technique used to amplify a specific region of DNA. It involves repeated cycles of heating and cooling of the DNA sample in the presence of DNA polymerase, primers that flank the target region, and dNTPs. Each cycle doubles the amount of target DNA, exponentially amplifying the target region up to millions of copies. PCR is widely used in medical research, forensics, and other applications to generate numerous copies of a specific DNA segment.DNA replication-in-eukaryotes
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LAMP PCR is a nucleic acid amplification technique that uses a DNA polymerase with strand displacement activity. It requires 4-6 designed primers that recognize 6 distinct regions on the target gene. Amplification occurs isothermally without temperature cycling. It produces stem-loop DNA structures with dumbbell ends that are amplified quickly using loop primers. LAMP has advantages over PCR like higher specificity, greater detection limit, simpler operation without expensive equipment, and ability to use crude samples. It is a sensitive, specific and cost-effective technique used for diagnosing infectious diseases.DNA Stuctre and function 
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Dna analysis can be used for the identification of carriers of hereditary disorders.
For prenatal diagnosis of serious genetic conditions yearly diagnosis before the onset of symptoms is done MOLECULAR DIAGNOSIS OF GENETIC DISEASE
CYSTIC FIBROSIS:
Cystic fibrosis is a genetic disease that affect mostly lungs and also the pancreas.
Screening test:
It is a complex process
Large number of genetic alterations have to be done.
For eg : It is the one of the most common lethal autosomal recessive disorder in Europe.
It is caused by mutations to cystic fibrosis transmembrane conductance regulator(CFTR) gene .
Screening individuals who may be at risk for cystic fibrosis for 500 different mutations is a daunting task.
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SICKLE CELL ANEMIA:
It is a disorder where red blood cells become rigid and sticky and are shaped like sickle.
This irregularly shaped cells stucks in small blood vessels which can slow and block the blood flow and oxygen to all the parts of the body.
Theres no cure for sickle cell anemia.
Screening for sickle cell anemia:
SCA is a genetic disease that is the result of a single nucleotide change in the codon for the sixth aminoacid of the 硫- chain of the hemoglobin molecule.
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Polymerase Chain Reaction (PCR) is a technique used to amplify small amounts of DNA sequences. It involves repeated cycles of heating and cooling of the DNA sample to denature and replicate the target DNA. Each cycle doubles the amount of target DNA, exponentially increasing its quantity for analysis. PCR uses primers, DNA polymerase, and dNTPs to selectively amplify the target DNA sequence. It has revolutionized molecular biology and is widely used for DNA cloning, detection of genetic diseases and mutations, forensic analysis, and more.Eukaryotic transcription
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Transcription in eukaryotes: A brief view
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In Eukaryotic transcription, synthesis of RNA occurs in the 35 direction. The 3 end is more reactive due to the hydroxide group. 5 end containing phosphate groups meanwhile, is not very reactive when it comes to adding new nucleotides. In Eukaryotes, the whole genome is not transcribed at once. Only a part of the genome is transcribed which also acts as the first, principle stage of genetic regulation.
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RNA Polymerase I: This produces rRNA (23S, 5.8S, and 18S) which are the major components in a ribosome. This also produces pre-rRNA in yeasts.
RNA Polymerase II: Helps in the production of mRNA (messenger RNA), snRNA (small, nuclear RNA), miRNA. This is the most studied type and requires several transcription factors for its binding
RNA Polymerase III: This synthesizes tRNA (transfer RNA), 5S rRNA and other small RNAs required in the cytosol and nucleus.
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Transcription is an elaborate process which cells use to copy the genetic information stored in DNA into RNA. This pre-RNA is modified into mRNA before being transcribed to proteins. Transcription is the first step to utilizing the genetic information in a cell. Both Eukaryotes and Prokaryotes employ this process with the basic phases remaining the same. However eukaryotic transcription is more complex indicating the changes transcription has undergone towards perfection during evolution. Ngs ppt
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The polymerase chain reaction (PCR) is an in vitro technique used to amplify a specific region of DNA. It involves repeated cycles of heating and cooling of the DNA sample in the presence of DNA polymerase, primers that flank the target region, and dNTPs. Each cycle doubles the amount of target DNA, exponentially amplifying the target region up to millions of copies. PCR is widely used in medical research, forensics, and other applications to generate numerous copies of a specific DNA segment.DNA replication-in-eukaryotes
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DEFINITION:
Dna analysis can be used for the identification of carriers of hereditary disorders.
For prenatal diagnosis of serious genetic conditions yearly diagnosis before the onset of symptoms is done MOLECULAR DIAGNOSIS OF GENETIC DISEASE
CYSTIC FIBROSIS:
Cystic fibrosis is a genetic disease that affect mostly lungs and also the pancreas.
Screening test:
It is a complex process
Large number of genetic alterations have to be done.
For eg : It is the one of the most common lethal autosomal recessive disorder in Europe.
It is caused by mutations to cystic fibrosis transmembrane conductance regulator(CFTR) gene .
Screening individuals who may be at risk for cystic fibrosis for 500 different mutations is a daunting task.
Diagnosis test that screen for a large number of mutations of a single gene in one assay being developed.
SICKLE CELL ANEMIA:
It is a disorder where red blood cells become rigid and sticky and are shaped like sickle.
This irregularly shaped cells stucks in small blood vessels which can slow and block the blood flow and oxygen to all the parts of the body.
Theres no cure for sickle cell anemia.
Screening for sickle cell anemia:
SCA is a genetic disease that is the result of a single nucleotide change in the codon for the sixth aminoacid of the 硫- chain of the hemoglobin molecule.
The anemia is caused by the inability of the mutated hemoglobin to carry sufficient oxygen.
Target probe hybridasation is done.
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This document summarizes the synthesis and structure of mRNA. It discusses that mRNA is transcribed from DNA in the nucleus and then undergoes processing before being transported to the cytoplasm for protein synthesis. The structure of mRNA includes a 5' cap, coding region, untranslated regions, and a poly-A tail. Transcription is carried out by RNA polymerase II and involves initiation, elongation, and termination. The primary transcript then undergoes processing where introns are removed and exons are joined through splicing.Polymerase chain reaction (PCR)
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The polymerase chain reaction (PCR) is an in vitro technique used to amplify specific DNA sequences. It involves repeated cycles of heating and cooling of the DNA sample to separate the DNA strands and allow for replication by DNA polymerase. Three main steps in each cycle are denaturation to separate the strands, annealing of primers to the target DNA, and extension of the new strands. PCR can generate millions of copies of target DNA from a very small sample and is used for a variety of applications including disease diagnosis, DNA fingerprinting, and molecular cloning.Polymerase Chain Reaction
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The Polymerase Chain Reaction (PCR) is a technique that allows for the amplification of specific DNA sequences. It involves cycling between heating and cooling steps to denature, anneal primers to, and extend DNA. This allows a small amount of DNA to be exponentially replicated, enabling applications like disease diagnosis, genetic identification, and DNA analysis. PCR requires DNA, primers, DNA polymerase, nucleotides, and thermal cycling to replicate the target DNA sequence.DNA replication in prokaryotes
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The document summarizes the process of DNA replication in prokaryotic cells like E. coli. It describes the three main stages: initiation at the Ori C origin site involving DnaA and DnaB proteins, elongation where the leading strand is synthesized continuously and the lagging strand requires RNA primers, and termination when the replication forks meet specific termination sites and DnaB is released. DNA replication allows faithful copying of the parental DNA to produce two identical progeny DNA molecules.Dna sequencing methods
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DNA sequencing techniques have evolved over time. The dideoxy method developed by Sanger is useful for sequencing short DNA fragments of 500-750bp by terminating the chain with ddNTPs. Whole genome sequencing became possible using shotgun sequencing, which breaks the genome into random fragments that are sequenced and then reassembled. More recently, pyrosequencing was developed for sequencing by synthesis and allows accurate, parallel, and automated sequencing without the need for gel electrophoresis.Eukaryotes replication
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油
DNA replication is the process by which DNA copies itself to produce identical daughter molecules. It occurs through a semi-conservative process in both prokaryotes and eukaryotes. In eukaryotes, replication is a complex multi-stage process involving initiation, formation of a pre-replication complex, conversion to an initiation complex, elongation via leading and lagging strand synthesis, and termination. Key proteins involved include ORC, Cdc6, Cdt1, MCM complexes, Cdc45, GINS, DNA polymerases, primase, ligase and topoisomerases. Replication occurs with high fidelity and is tightly regulated to prevent re-replication.Telomerase
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Telomeres are repetitive nucleotide sequences located at the ends of chromosomes that protect the ends from deterioration or fusion with other chromosomes. They shorten each time a cell divides until a length is reached that triggers cell senescence or death. The enzyme telomerase maintains telomere length by adding telomeric repeats to chromosome ends using its own RNA component as a template. Telomerase is active in germ cells, stem cells, and cancer cells, allowing indefinite cell division, but not in most somatic cells, limiting their replicative lifespan.Polymerase Chain Reaction(PCR)
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油
Polymerase Chain Reaction (PCR) is a technique used to amplify small amounts of DNA sequences. It involves repeated cycles of heating and cooling of the DNA sample to denature and replicate the target DNA. Each cycle doubles the amount of target DNA, exponentially increasing its quantity for analysis. PCR uses primers, DNA polymerase, and dNTPs to selectively amplify the target DNA sequence. It has revolutionized molecular biology and is widely used for DNA cloning, detection of genetic diseases and mutations, forensic analysis, and more.Eukaryotic transcription
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油
Transcription in eukaryotes: A brief view
Transcription is the process by which single stranded RNA is synthesized by double stranded DNA. Transcription in eukaryotes and prokaryotes has many similarities while at the same time both showing their individual characteristics due to the differences in organization. RNA Polymerase (RNAP or RNA Pol) is different in prokaryotes and eukaryotes. Coupled transcription is seen in prokaryotes but not in Eukaryotes. In eukaryotes the pre-RNA should be spliced first to be translated.
In Eukaryotic transcription, synthesis of RNA occurs in the 35 direction. The 3 end is more reactive due to the hydroxide group. 5 end containing phosphate groups meanwhile, is not very reactive when it comes to adding new nucleotides. In Eukaryotes, the whole genome is not transcribed at once. Only a part of the genome is transcribed which also acts as the first, principle stage of genetic regulation.
Eukaryotes have five nuclear polymerases:
RNA Polymerase I: This produces rRNA (23S, 5.8S, and 18S) which are the major components in a ribosome. This also produces pre-rRNA in yeasts.
RNA Polymerase II: Helps in the production of mRNA (messenger RNA), snRNA (small, nuclear RNA), miRNA. This is the most studied type and requires several transcription factors for its binding
RNA Polymerase III: This synthesizes tRNA (transfer RNA), 5S rRNA and other small RNAs required in the cytosol and nucleus.
RNA Polymerase IV: Synthesizes siRNA (small interfering RNA) in plants.
RNA Polymerase V: This is the least studied polymerase and synthesizes siRNA-directed heterochromatin in plants.
Eukaryotic transcription can be broadly divided into 4 stages:
Pre-Initiation
Initiation
Elongation
Termination
Transcription is an elaborate process which cells use to copy the genetic information stored in DNA into RNA. This pre-RNA is modified into mRNA before being transcribed to proteins. Transcription is the first step to utilizing the genetic information in a cell. Both Eukaryotes and Prokaryotes employ this process with the basic phases remaining the same. However eukaryotic transcription is more complex indicating the changes transcription has undergone towards perfection during evolution. Ngs ppt
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This document provides an overview of next generation sequencing (NGS) technologies. It discusses the history and evolution of DNA sequencing, from early manual methods developed by Sanger to modern high-throughput NGS approaches. Key NGS methods described include Illumina sequencing by synthesis, Ion Torrent semiconductor sequencing, 454 pyrosequencing, and SOLiD ligation sequencing. Compared to Sanger, NGS allows massively parallel sequencing of many samples at lower cost and higher throughput. While NGS has advanced biological research, each method still has advantages and limitations related to read length, accuracy, and cost.Similar to PCR --Polymerase Chain Reaction (20)
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This document provides an overview of polymerase chain reaction (PCR) and electrophoresis techniques. It discusses the basic principles and practical applications of PCR, including DNA amplification, temperature cycling, and primer design. The document also reviews the history and development of PCR by Kary Mullis. It explains the three main steps of the PCR process - denaturation, annealing and extension. Additionally, it discusses various PCR techniques like real-time PCR and reverse transcription PCR. The document provides details on materials, reagents and procedures for performing PCR. It also outlines applications of PCR and electrophoresis in various fields like medicine, forensics and research.PCR
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Polymerase Chin Reaction is a technique that takes specific sequences of DNA of small and amplifies it to be used for further testing.
it is also said to be as the Invitro Technique.We have seen an photocopy machine in an office, by which we can copy several pages. So, is the PCR machine in a molecular biology laboratory.
PCR is DNA raplication ina test tube.
Dr Kary Mullis developed PCR.
To amplify lot of double stranded DNA molecules with same size and sequence by enzymatic method and cycling condition.
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PCR is a technique that amplifies a specific DNA sequence from a small sample. It involves denaturing DNA, annealing primers to the DNA, and extending the DNA strands using a DNA polymerase enzyme in repeated cycles. This process amplifies the target DNA sequence exponentially, allowing it to be detected and analyzed. PCR has many applications including disease diagnosis, genetic testing, forensics, and molecular biology research.PCR techenology in fisheries
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The polymerase chain reaction (PCR) is a technique for amplifying DNA segments. It involves denaturing DNA, annealing primers to the single strands, and extending the primers to replicate the DNA segment. Kary Mullis developed PCR in 1983 and won the Nobel Prize for it. PCR consists of cycles of heating and cooling DNA which allows exponential amplification of specific DNA regions defined by primer binding sites. It has many applications in research, forensics, medicine and more.
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Polymerase chain reaction (PCR) is a technique used to amplify DNA sequences. It was developed by Kary Mullis in 1983 and has become a common technique used in medical and biological research. Mullis received the Nobel Prize in Chemistry in 1993 for his work on PCR. PCR involves denaturing DNA, annealing primers to the DNA, and extending the primers to replicate the DNA in multiple cycles, exponentially amplifying any specific DNA region. There are various types of PCR including conventional PCR, multiplex PCR, nested PCR, quantitative PCR, and real-time PCR which have various applications such as disease diagnosis, DNA sequencing, and phylogeny analysis.Recently uploaded (20)
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PCR --Polymerase Chain Reaction
- 1. PCR---Polymerase Chain Reaction M Adnan Qadar Your credits at the end of slideshow; Motivate Encourage & Recommend us Remember to credit us by clicking on the last slide to produce quality contents.
- 2. Polymerase Chain Reaction (PCR) Q. What is PCR? Ans. A technique of Molecular Biology that amplifies a specific segment of DNA to form its thousands to millions of copies in efficient way. Q. What are the benefits of PCR? Ans. Following are the applications of PCR in daily life; 1. Increasing the efficiency of DNA/gene of interest copying 2. A source of gene library formation 3. Efficiency in Forensic identification of criminals 4. Resolving the disputed paternity cases 5. Clinical diagnostics of pathogens
- 3. PCR---History Development: 1983 Kary B. Mullis developed PCR 1989 Molecule of the year was denoted to Taq Polymerase 1993 Mullis shared the Noble Prize in chemistry for PCR technology About Mullis: A U.S. Scientist Born on Dec 28, 1944 North Carolina Died Aug 07, 2019 California Fish don't know much about water, and people didn't know much about air K. B. Mullis
- 5. PCR---Requirements OR Ingredients Following are the requirements to carry out the PCR; 1. DNA/Piece of DNA (To be Copied) 2. Primers (for Identification of starting point at DNA strands) 3. Taq polymerase (works normal till 95属C) 4. dNTPs (dATP, dTTP, dCTP, dGTP) 5. Buffers (Stabilizing reaction mixture during PCR) 6. MgCl2 (Cofactor for Taq Polymerase) 7. Thermocycler (Machine to carry out the PCR)
- 6. PCR---PROCESS 1. Mixing all the ingredients in a DNA tube (in recommended amounts) 2. Programming the DNA reaction mixture into the thermocycler Function of Thermocycler: A Machine that is Programmed to heat and cool the mixture within short period of time (seconds-minutes) over a number of cycles
- 7. Steps of---PCR: Following are the main steps in PCR; 1. Denaturation 2. Annealing 3. Extension PCR---PROCESS
- 8. Denaturation: 1. Thermocycler shall heat the Mixture (Up to 96属C) 2. This leads to breaking of nucleotides bonds 3. Separation of double strands of DNA Annealing: 1. Thermocycler cools the Mixture (Up to 54属C ) 2. This enables the Primers to Identify & attach the DNA sites for formation of new strands PCR---PROCESS
- 9. Extension: 1. Attachment of Taq Polymerase at 3卒 ends of primer 2. Thermocycler will increase the temperature (Up to 96属C) 3. Taq Polymerase shall extend/polymerize the new strands of DNA by picking the respective dNTPs from the reaction mixture PCR---PROCESS
- 10. Final hold---Conclusion 1. Finally the newly formed DNA hold at 4属C 2. By this process one can prepare a number of abundant copies of required DNA within minutes of cycles
- 11. ANIMATION---CLICK TO CRDIT ANIMATION CLICK TO CREDIT THANKS FOR WATCHING