Suche senden
Hochladen
Polymerase chain reaction medical school
•
2 gefällt mir
•
436 views
Ravi Kiran
Folgen
BIOCHEMISTRY
Weniger lesen
Mehr lesen
Gesundheit & Medizin
Melden
Teilen
Melden
Teilen
1 von 32
Jetzt herunterladen
Downloaden Sie, um offline zu lesen
Empfohlen
Polymerase chain reaction
Polymerase chain reaction
Virendra Baghel
M sc2
M sc2
DESH BANDHU GANGWAR
IFPAC 2013 Baltimore
IFPAC 2013 Baltimore
dominev
PCR
PCR
tarek aziz
Syed pcr
Syed pcr
syed fayazuddin
Amplification of gene using PCR
Amplification of gene using PCR
Vidya Kalaivani Rajkumar
PhD Poster - UKEMS Conference 2008
PhD Poster - UKEMS Conference 2008
Donna Johnson
Gene 151_119 (1994) [SDM of dsDNA]
Gene 151_119 (1994) [SDM of dsDNA]
Michael Weiner
Weitere ähnliche Inhalte
Was ist angesagt?
Polymerase Chain Reaction
Polymerase Chain Reaction
JasleenSaini10
12.18.09 Theis Lab Group Presentation
12.18.09 Theis Lab Group Presentation
guestd626c0
PCR BASED MOLECULARMARKERS
PCR BASED MOLECULARMARKERS
ANANDALEKSHMIL
Polymerase Chain Reaction(PCR)
Polymerase Chain Reaction(PCR)
parnavi kadam
Izzatullah presentation 11824
Izzatullah presentation 11824
UsmanAli996
Pcr Presentation
Pcr Presentation
Tasmina Susmi
An Investigation Of The Rigor Of Interpretation Rules
An Investigation Of The Rigor Of Interpretation Rules
Nick Brown
Alberto Kornblihtt-Enfermedades raras de la piel
Alberto Kornblihtt-Enfermedades raras de la piel
Fundación Ramón Areces
Polymerase Chain Reaction
Polymerase Chain Reaction
sara_abudahab
Lab talk 201109 radioligand assay for validating in slico predicted rel1 anta...
Lab talk 201109 radioligand assay for validating in slico predicted rel1 anta...
Laurence Dawkins-Hall
RNA editing as a drug target in tryp. development of a high throughput sceeni...
RNA editing as a drug target in tryp. development of a high throughput sceeni...
Laurence Dawkins-Hall
Polymerase Chain Reaction Practical
Polymerase Chain Reaction Practical
Sabahat Ali
A rapid dna assembling strategy mediated by direct full length
A rapid dna assembling strategy mediated by direct full length
juansebastianpz
Polymerase chain reaction Pranav
Polymerase chain reaction Pranav
Pranav S
Nested PCR,Fusion primers, Integrated PCR
Nested PCR,Fusion primers, Integrated PCR
Sani Jwel
PCR and RT-PCR
PCR and RT-PCR
Raghavendra institute of pharmaceutical education and research .
POLYMERASE CHAIN REACTION
POLYMERASE CHAIN REACTION
ankit
19_21Translation
19_21Translation
Karen Lewis
Polymerase chain reaction(pcr)
Polymerase chain reaction(pcr)
abrishiya
IRJET- Subcellular Localization of Transmembrane E-cadherin-GFP Fusion Pr...
IRJET- Subcellular Localization of Transmembrane E-cadherin-GFP Fusion Pr...
IRJET Journal
Was ist angesagt?
(20)
Polymerase Chain Reaction
Polymerase Chain Reaction
12.18.09 Theis Lab Group Presentation
12.18.09 Theis Lab Group Presentation
PCR BASED MOLECULARMARKERS
PCR BASED MOLECULARMARKERS
Polymerase Chain Reaction(PCR)
Polymerase Chain Reaction(PCR)
Izzatullah presentation 11824
Izzatullah presentation 11824
Pcr Presentation
Pcr Presentation
An Investigation Of The Rigor Of Interpretation Rules
An Investigation Of The Rigor Of Interpretation Rules
Alberto Kornblihtt-Enfermedades raras de la piel
Alberto Kornblihtt-Enfermedades raras de la piel
Polymerase Chain Reaction
Polymerase Chain Reaction
Lab talk 201109 radioligand assay for validating in slico predicted rel1 anta...
Lab talk 201109 radioligand assay for validating in slico predicted rel1 anta...
RNA editing as a drug target in tryp. development of a high throughput sceeni...
RNA editing as a drug target in tryp. development of a high throughput sceeni...
Polymerase Chain Reaction Practical
Polymerase Chain Reaction Practical
A rapid dna assembling strategy mediated by direct full length
A rapid dna assembling strategy mediated by direct full length
Polymerase chain reaction Pranav
Polymerase chain reaction Pranav
Nested PCR,Fusion primers, Integrated PCR
Nested PCR,Fusion primers, Integrated PCR
PCR and RT-PCR
PCR and RT-PCR
POLYMERASE CHAIN REACTION
POLYMERASE CHAIN REACTION
19_21Translation
19_21Translation
Polymerase chain reaction(pcr)
Polymerase chain reaction(pcr)
IRJET- Subcellular Localization of Transmembrane E-cadherin-GFP Fusion Pr...
IRJET- Subcellular Localization of Transmembrane E-cadherin-GFP Fusion Pr...
Andere mochten auch
"New Realities in Asian Markets for Ireland:The Success of Origin Green" Aida...
"New Realities in Asian Markets for Ireland:The Success of Origin Green" Aida...
Asia Matters
Milestones Reached, More to Come: Challenges to Overcome in Concluding the Ja...
Milestones Reached, More to Come: Challenges to Overcome in Concluding the Ja...
Asia Matters
Constructivism
Constructivism
Cesar C. Navarrete
Eileen Sharpe, Director of Growth Markets, IDA Ireland, Asia Business Week Du...
Eileen Sharpe, Director of Growth Markets, IDA Ireland, Asia Business Week Du...
Asia Matters
Chemistry of nucleotides
Chemistry of nucleotides
Ravi Kiran
Rbpphhyper 131204183150-phpapp02
Rbpphhyper 131204183150-phpapp02
Jinalyn Calma
eFax Corporate Secure Faxing Online Datasheet
eFax Corporate Secure Faxing Online Datasheet
eFax Corporate®
Lake tahoe basin
Lake tahoe basin
Perry Quinn
Future of marketing the cloud john fearon, drop mysite
Future of marketing the cloud john fearon, drop mysite
Binexa Technology
Middle School Game Development Research
Middle School Game Development Research
Cesar C. Navarrete
“Delivering Real Education Partnership between Asia and Ireland” Professor Ci...
“Delivering Real Education Partnership between Asia and Ireland” Professor Ci...
Asia Matters
"Talent for Innovation: The Role of Universities." Ned Costello
"Talent for Innovation: The Role of Universities." Ned Costello
Asia Matters
"Ireland's Leadership Role in Creating Global Value Chains in the AgriFood Se...
"Ireland's Leadership Role in Creating Global Value Chains in the AgriFood Se...
Asia Matters
Vision planet
Vision planet
Igor Vasilev
Guide To Successful Affiliate Marketing
Guide To Successful Affiliate Marketing
shawn300
Prof. David FitzPatrick, Provost, Beijing-Dublin International School
Prof. David FitzPatrick, Provost, Beijing-Dublin International School
Asia Matters
"Attracting Blue Chip Chinese Firms to Set Up a European base in Ireland" Eil...
"Attracting Blue Chip Chinese Firms to Set Up a European base in Ireland" Eil...
Asia Matters
Andere mochten auch
(17)
"New Realities in Asian Markets for Ireland:The Success of Origin Green" Aida...
"New Realities in Asian Markets for Ireland:The Success of Origin Green" Aida...
Milestones Reached, More to Come: Challenges to Overcome in Concluding the Ja...
Milestones Reached, More to Come: Challenges to Overcome in Concluding the Ja...
Constructivism
Constructivism
Eileen Sharpe, Director of Growth Markets, IDA Ireland, Asia Business Week Du...
Eileen Sharpe, Director of Growth Markets, IDA Ireland, Asia Business Week Du...
Chemistry of nucleotides
Chemistry of nucleotides
Rbpphhyper 131204183150-phpapp02
Rbpphhyper 131204183150-phpapp02
eFax Corporate Secure Faxing Online Datasheet
eFax Corporate Secure Faxing Online Datasheet
Lake tahoe basin
Lake tahoe basin
Future of marketing the cloud john fearon, drop mysite
Future of marketing the cloud john fearon, drop mysite
Middle School Game Development Research
Middle School Game Development Research
“Delivering Real Education Partnership between Asia and Ireland” Professor Ci...
“Delivering Real Education Partnership between Asia and Ireland” Professor Ci...
"Talent for Innovation: The Role of Universities." Ned Costello
"Talent for Innovation: The Role of Universities." Ned Costello
"Ireland's Leadership Role in Creating Global Value Chains in the AgriFood Se...
"Ireland's Leadership Role in Creating Global Value Chains in the AgriFood Se...
Vision planet
Vision planet
Guide To Successful Affiliate Marketing
Guide To Successful Affiliate Marketing
Prof. David FitzPatrick, Provost, Beijing-Dublin International School
Prof. David FitzPatrick, Provost, Beijing-Dublin International School
"Attracting Blue Chip Chinese Firms to Set Up a European base in Ireland" Eil...
"Attracting Blue Chip Chinese Firms to Set Up a European base in Ireland" Eil...
Ähnlich wie Polymerase chain reaction medical school
PCR.ppt
PCR.ppt
Dr. R. DINESHKUMAR
PCR.ppt
PCR.ppt
Dr. R. DINESHKUMAR
Polymerase Chain Reaction
Polymerase Chain Reaction
DrSushilKumarSoni
Amplificationtechniquesinmicrobiology 090609205257-phpapp02
Amplificationtechniquesinmicrobiology 090609205257-phpapp02
minhdaovan
Dwd pcr
Dwd pcr
deepak deshkar
Polymerase chain reaction & electrophoresis
Polymerase chain reaction & electrophoresis
Bahauddin Zakariya University lahore
PCR lecture.ppt
PCR lecture.ppt
NoorKhan428102
Polymerase chain reaction (pcr)
Polymerase chain reaction (pcr)
RameshPandi4
Polymerase chain reaction principles and practice
Polymerase chain reaction principles and practice
subramaniam sethupathy
Pcr
Pcr
Dr Alok Bharti
PCR_2017.pptx
PCR_2017.pptx
lalvarezmex
Pcr 29 07-2011 final
Pcr 29 07-2011 final
Dr Praveen kumar . V Doddamani
POLYMERASE CHAIN REACTION(PCR)-subina sunar.pptx
POLYMERASE CHAIN REACTION(PCR)-subina sunar.pptx
SubinaSunar
PCR and its types
PCR and its types
sujathar23
Pcr and its application
Pcr and its application
christanantony
Technique of polymerase chain reaction (pcr) experimental biotechnology
Technique of polymerase chain reaction (pcr) experimental biotechnology
St.Xavier's College , Palayamkottai - 627 002
PCR Methods and applications
PCR Methods and applications
Behzad Milani
Polymerase chain reaction
Polymerase chain reaction
Siddhartha Roy
PCR
PCR
Hemanth Muckatira
PCR and RT PCR.pptx
PCR and RT PCR.pptx
RajkumarYadav587017
Ähnlich wie Polymerase chain reaction medical school
(20)
PCR.ppt
PCR.ppt
PCR.ppt
PCR.ppt
Polymerase Chain Reaction
Polymerase Chain Reaction
Amplificationtechniquesinmicrobiology 090609205257-phpapp02
Amplificationtechniquesinmicrobiology 090609205257-phpapp02
Dwd pcr
Dwd pcr
Polymerase chain reaction & electrophoresis
Polymerase chain reaction & electrophoresis
PCR lecture.ppt
PCR lecture.ppt
Polymerase chain reaction (pcr)
Polymerase chain reaction (pcr)
Polymerase chain reaction principles and practice
Polymerase chain reaction principles and practice
Pcr
Pcr
PCR_2017.pptx
PCR_2017.pptx
Pcr 29 07-2011 final
Pcr 29 07-2011 final
POLYMERASE CHAIN REACTION(PCR)-subina sunar.pptx
POLYMERASE CHAIN REACTION(PCR)-subina sunar.pptx
PCR and its types
PCR and its types
Pcr and its application
Pcr and its application
Technique of polymerase chain reaction (pcr) experimental biotechnology
Technique of polymerase chain reaction (pcr) experimental biotechnology
PCR Methods and applications
PCR Methods and applications
Polymerase chain reaction
Polymerase chain reaction
PCR
PCR
PCR and RT PCR.pptx
PCR and RT PCR.pptx
Mehr von Ravi Kiran
Heme Structure. synthesis and porphyrias
Heme Structure. synthesis and porphyrias
Ravi Kiran
Steroid hormones
Steroid hormones
Ravi Kiran
Structure of DNA
Structure of DNA
Ravi Kiran
Chemistry of nucleotides
Chemistry of nucleotides
Ravi Kiran
Chemistry of lipids phospholipids
Chemistry of lipids phospholipids
Ravi Kiran
Chemistry of chemistry of lipids
Chemistry of chemistry of lipids
Ravi Kiran
Chemistry of lipids. Part 1
Chemistry of lipids. Part 1
Ravi Kiran
Chemistry of carbohydrates polysaccharides part 3 B heteroglycans
Chemistry of carbohydrates polysaccharides part 3 B heteroglycans
Ravi Kiran
Chemistry of carbohydrates polysaccharides part -3 homoglycans
Chemistry of carbohydrates polysaccharides part -3 homoglycans
Ravi Kiran
Chemistry of carbohydrates disaccharides
Chemistry of carbohydrates disaccharides
Ravi Kiran
Chemistry of carbohydrates
Chemistry of carbohydrates
Ravi Kiran
Membrane structure and transport for medical school
Membrane structure and transport for medical school
Ravi Kiran
Digestion and absorption of proteins for Medical School
Digestion and absorption of proteins for Medical School
Ravi Kiran
Digestion and absorption of carbohydrates for Medical School
Digestion and absorption of carbohydrates for Medical School
Ravi Kiran
Lipid digestion and absorption for medical school
Lipid digestion and absorption for medical school
Ravi Kiran
The cell for medical school
The cell for medical school
Ravi Kiran
Gluconeogenesis for medical school
Gluconeogenesis for medical school
Ravi Kiran
Structure of RNA. To teach Ist year medical students.
Structure of RNA. To teach Ist year medical students.
Ravi Kiran
Structure of DNA for medical school
Structure of DNA for medical school
Ravi Kiran
Oxidative phosphorylation for medical school
Oxidative phosphorylation for medical school
Ravi Kiran
Mehr von Ravi Kiran
(20)
Heme Structure. synthesis and porphyrias
Heme Structure. synthesis and porphyrias
Steroid hormones
Steroid hormones
Structure of DNA
Structure of DNA
Chemistry of nucleotides
Chemistry of nucleotides
Chemistry of lipids phospholipids
Chemistry of lipids phospholipids
Chemistry of chemistry of lipids
Chemistry of chemistry of lipids
Chemistry of lipids. Part 1
Chemistry of lipids. Part 1
Chemistry of carbohydrates polysaccharides part 3 B heteroglycans
Chemistry of carbohydrates polysaccharides part 3 B heteroglycans
Chemistry of carbohydrates polysaccharides part -3 homoglycans
Chemistry of carbohydrates polysaccharides part -3 homoglycans
Chemistry of carbohydrates disaccharides
Chemistry of carbohydrates disaccharides
Chemistry of carbohydrates
Chemistry of carbohydrates
Membrane structure and transport for medical school
Membrane structure and transport for medical school
Digestion and absorption of proteins for Medical School
Digestion and absorption of proteins for Medical School
Digestion and absorption of carbohydrates for Medical School
Digestion and absorption of carbohydrates for Medical School
Lipid digestion and absorption for medical school
Lipid digestion and absorption for medical school
The cell for medical school
The cell for medical school
Gluconeogenesis for medical school
Gluconeogenesis for medical school
Structure of RNA. To teach Ist year medical students.
Structure of RNA. To teach Ist year medical students.
Structure of DNA for medical school
Structure of DNA for medical school
Oxidative phosphorylation for medical school
Oxidative phosphorylation for medical school
Kürzlich hochgeladen
Rheumatoid arthritis Part 1, case based approach with application of the late...
Rheumatoid arthritis Part 1, case based approach with application of the late...
Internal medicine department, faculty of Medicine Beni-Suef University Egypt
GOUT UPDATE AHMED YEHIA 2024, case based approach with application of the lat...
GOUT UPDATE AHMED YEHIA 2024, case based approach with application of the lat...
Internal medicine department, faculty of Medicine Beni-Suef University Egypt
AI in Action: Elevating Patient Insights
AI in Action: Elevating Patient Insights
Emily Kunka, MS, CCRP
Adenomyosis or Fibroid- making right diagnosis
Adenomyosis or Fibroid- making right diagnosis
Sujoy Dasgupta
Male Infertility, Antioxidants and Beyond
Male Infertility, Antioxidants and Beyond
Sujoy Dasgupta
pA2 value, Schild plot and pD2 values- applications in pharmacology
pA2 value, Schild plot and pD2 values- applications in pharmacology
DeepakDaniel9
SGK RỐI LOẠN TOAN KIỀM ĐHYHN RẤT HAY VÀ ĐẶC SẮC.pdf
SGK RỐI LOẠN TOAN KIỀM ĐHYHN RẤT HAY VÀ ĐẶC SẮC.pdf
HongBiThi1
Employability skills, work experience presentation
Employability skills, work experience presentation
marwaahmad357
Arthroscopic Surgery in Indore : A Minimally Invasive Guide to Joint Health
Arthroscopic Surgery in Indore : A Minimally Invasive Guide to Joint Health
Gokuldas Hospital
Transport across cell membrane (passive, active, vesicular)
Transport across cell membrane (passive, active, vesicular)
MedicoseAcademics
blood bank management system project report
blood bank management system project report
NARMADAPETROLEUMGAS
American College of physicians ACP high value care recommendations in rheumat...
American College of physicians ACP high value care recommendations in rheumat...
Internal medicine department, faculty of Medicine Beni-Suef University Egypt
Understanding AIDS:a roadmap to well ess
Understanding AIDS:a roadmap to well ess
AbhishekKumar524514
PAIN/CLASSIFICATION AND MANAGEMENT OF PAIN.pdf
PAIN/CLASSIFICATION AND MANAGEMENT OF PAIN.pdf
Dolisha Warbi
SGK ĐIỆN GIẬT ĐHYHN RẤT LÀ HAY TUYỆT VỜI.pdf
SGK ĐIỆN GIẬT ĐHYHN RẤT LÀ HAY TUYỆT VỜI.pdf
HongBiThi1
Generative AI in Health Care a scoping review and a persoanl experience.
Generative AI in Health Care a scoping review and a persoanl experience.
Vaikunthan Rajaratnam
Immune labs basics part 1 acute phase reactants ESR, CRP Ahmed Yehia Ismaeel,...
Immune labs basics part 1 acute phase reactants ESR, CRP Ahmed Yehia Ismaeel,...
Internal medicine department, faculty of Medicine Beni-Suef University Egypt
BENIGN BREAST DISEASE
BENIGN BREAST DISEASE
Mamatha Lakka
ayurvedic formulations herbal drug technologyppt
ayurvedic formulations herbal drug technologyppt
Pradnya Wadekar
Neurological history taking (2024) .
Neurological history taking (2024) .
Mohamed Rizk Khodair
Kürzlich hochgeladen
(20)
Rheumatoid arthritis Part 1, case based approach with application of the late...
Rheumatoid arthritis Part 1, case based approach with application of the late...
GOUT UPDATE AHMED YEHIA 2024, case based approach with application of the lat...
GOUT UPDATE AHMED YEHIA 2024, case based approach with application of the lat...
AI in Action: Elevating Patient Insights
AI in Action: Elevating Patient Insights
Adenomyosis or Fibroid- making right diagnosis
Adenomyosis or Fibroid- making right diagnosis
Male Infertility, Antioxidants and Beyond
Male Infertility, Antioxidants and Beyond
pA2 value, Schild plot and pD2 values- applications in pharmacology
pA2 value, Schild plot and pD2 values- applications in pharmacology
SGK RỐI LOẠN TOAN KIỀM ĐHYHN RẤT HAY VÀ ĐẶC SẮC.pdf
SGK RỐI LOẠN TOAN KIỀM ĐHYHN RẤT HAY VÀ ĐẶC SẮC.pdf
Employability skills, work experience presentation
Employability skills, work experience presentation
Arthroscopic Surgery in Indore : A Minimally Invasive Guide to Joint Health
Arthroscopic Surgery in Indore : A Minimally Invasive Guide to Joint Health
Transport across cell membrane (passive, active, vesicular)
Transport across cell membrane (passive, active, vesicular)
blood bank management system project report
blood bank management system project report
American College of physicians ACP high value care recommendations in rheumat...
American College of physicians ACP high value care recommendations in rheumat...
Understanding AIDS:a roadmap to well ess
Understanding AIDS:a roadmap to well ess
PAIN/CLASSIFICATION AND MANAGEMENT OF PAIN.pdf
PAIN/CLASSIFICATION AND MANAGEMENT OF PAIN.pdf
SGK ĐIỆN GIẬT ĐHYHN RẤT LÀ HAY TUYỆT VỜI.pdf
SGK ĐIỆN GIẬT ĐHYHN RẤT LÀ HAY TUYỆT VỜI.pdf
Generative AI in Health Care a scoping review and a persoanl experience.
Generative AI in Health Care a scoping review and a persoanl experience.
Immune labs basics part 1 acute phase reactants ESR, CRP Ahmed Yehia Ismaeel,...
Immune labs basics part 1 acute phase reactants ESR, CRP Ahmed Yehia Ismaeel,...
BENIGN BREAST DISEASE
BENIGN BREAST DISEASE
ayurvedic formulations herbal drug technologyppt
ayurvedic formulations herbal drug technologyppt
Neurological history taking (2024) .
Neurological history taking (2024) .
Polymerase chain reaction medical school
1.
©2001 Timothy G.
Standish V.S.RAVIKIRAN, MSc. Polymerase Chain Reaction
2.
V.S.RAVIKIRAN, MSc., Department of
Biochemistry, ASRAM Medical college, Eluru-534005.AP, India. vsravikiran2013@gmail.com
3.
©2001 Timothy G.
Standish V.S.RAVIKIRAN, MSc., Department of Biochemistry, ASRAM Medical college, Eluru-534005.AP, India. vsravikiran2013@gmail.com
4.
©2001 Timothy G.
Standish Polymerase ChainPolymerase Chain ReactionReaction V.S.RAVIKIRAN
5.
©2001 Timothy G.
Standish HistoryHistory The Polymerase Chain Reaction (PCR) was not a discovery, but rather an invention A special DNA polymerase (Taq) is used to make many copies of a short length of DNA (100-10,000 bp) defined by primers Kary Mullis, the inventor of PCR, was awarded the 1993 Nobel Prize in Chemistry
6.
©2001 Timothy G.
Standish What PCR Can DoWhat PCR Can Do PCR can be used to make many copies of any DNA that is supplied as a template Starting with one original copy an almost infinite number of copies can be made using PCR “Amplified” fragments of DNA can be sequenced, cloned, probed or sized using electrophoresis
7.
©2001 Timothy G.
Standish What PCR Can DoWhat PCR Can Do Defective genes can be amplified to diagnose any number of illnesses Genes from pathogens can be amplified to identify them (ie. HIV) Amplified fragments can act as genetic fingerprints
8.
©2001 Timothy G.
Standish How PCR WorksHow PCR Works PCR is an artificial way of doing DNA replication Instead of replicating all the DNA present, only a small segment is replicated, but this small segment is replicated many times
9.
©2001 Timothy G.
Standish How PCR WorksHow PCR Works As in replication, PCR involves: –Melting DNA –Priming –Polymerization
10.
©2001 Timothy G.
Standish Initiation - Forming theInitiation - Forming the Replication EyeReplication Eye 3’ 5’ 3’5’ 5’ 5’ 3’ 3’ Origin of Replication 5’ 3’ 3’ 5’ 5’ 3’ 5’ 5’ 5’ 3’ 3’ 3’
11.
©2001 Timothy G.
Standish Leading Strand Laging Strand 3’ 5’ 3’ 5’ Extension - The Replication ForkExtension - The Replication Fork 5’ 5’ 5’ 3’ 3’ 5’3’ 3’ 5’ Single strand binding proteins DNA Polymerase Okazaki fragment RNA Primers Primase 5’ 3’ 5’ Helicase
12.
©2001 Timothy G.
Standish Functions And TheirFunctions And Their Associated EnzymesAssociated Enzymes LigaseJoining nicks DNA PolymerasePolymerizing DNA PrimaseProviding primer EnzymeFunction Helicase SSB Proteins Topisomerase Melting DNA
13.
©2001 Timothy G.
Standish Components of a PCRComponents of a PCR ReactionReaction Buffer (containing Mg++ ) Template DNA 2 Primers that flank the fragment of DNA to be amplified dNTPs Taq DNA Polymerase (or another thermally stable DNA polymerase)
14.
©2001 Timothy G.
Standish PCRPCR Melting 94 o C Melting 94 o C Annealing Primers 50 o C Extension 72 o C Temperature 100 0 50 T i m e 30x 5’3’ 3’5’ 3’5’ 5’ 5’3’ 5’ 3’5’ 5’ 5’ 5’ 5’3’ 3’5’ 3’5’ 5’3’ 5’3’ 5’
15.
©2001 Timothy G.
Standish PCRPCRMelting 94 o C Temperature 100 0 50 T i m e 5’3’ 3’5’
16.
©2001 Timothy G.
Standish PCRPCRMelting 94 o C Temperature 100 0 50 T i m e 3’5’ 5’3’ Heat
17.
©2001 Timothy G.
Standish PCRPCRMelting 94 o C Annealing Primers 50 o C Extension 72 o CTemperature 100 0 50 T i m e 3’5’ 5’3’ 5’ 5’ Melting 94 o C
18.
©2001 Timothy G.
Standish PCRPCRMelting 94 o C Melting 94 o C Annealing Primers 50 o C Extension 72 o CTemperature 100 0 50 T i m e 30x 3’5’ 5’3’ Heat Heat 5’ 5’ 5’
19.
©2001 Timothy G.
Standish PCRPCRMelting 94 o C Melting 94 o C Annealing Primers 50 o C Extension 72 o CTemperature 100 0 50 T i m e 30x 3’5’ 5’3’ 5’ 5’ 5’ 5’ 5’ 5’
20.
©2001 Timothy G.
Standish PCRPCRMelting 94 o C Melting 94 o C Annealing Primers 50 o C Extension 72 o CTemperature 100 0 50 T i m e 30x 3’5’ 5’3’ 5’ 5’ 5’ 5’ 5’ 5’ Heat Heat
21.
©2001 Timothy G.
Standish PCRPCRMelting 94 o C Melting 94 o C Annealing Primers 50 o C Extension 72 o CTemperature 100 0 50 T i m e 30x 3’5’ 5’3’ 5’ 5’ 5’ 5’ 5’ 5’ 5’ 5’ 5’ 5’
22.
©2001 Timothy G.
Standish Fragments of defined length PCRPCRMelting 94 o C Melting 94 o C Annealing Primers 50 o C Extension 72 o CTemperature 100 0 50 T i m e 30x 3’5’ 5’3’ 5’ 5’ 5’ 5’ 5’ 5’ 5’ 5’ 5’ 5’
23.
©2001 Timothy G.
Standish DNA Between The Primers DoublesDNA Between The Primers Doubles With Each Thermal CycleWith Each Thermal Cycle 0 Cycles Number 1 3 8 2 4 1 2 4 16 5 32 6 64
24.
©2001 Timothy G.
Standish More Cycles = More DNAMore Cycles = More DNA Number of cycles 0 10 15 20 25 30 Size Marker
25.
©2001 Timothy G.
Standish Theoretical Yield Of PCRTheoretical Yield Of PCR Theoretical yield = 2n x y Where y = the starting number of copies and n = the number of thermal cycles = 107,374,182,400 If you start with 100 copies, how many copies are made in 30 cycles? 2n x y = 230 x 100 = 1,073,741,824 x 100
26.
©2001 Timothy G.
Standish How The Functions Of ReplicationHow The Functions Of Replication Are Achieved During PCRAre Achieved During PCR N/A as fragments are short Joining nicks Taq DNA PolymerasePolymerizing DNA Primers are added to the reaction mix Providing primer PCRFunction HeatMelting DNA
27.
©2001 Timothy G.
Standish 27 Type of Gene CyclerType of Gene Cycler Multi Block PCRStandard PCR Gradient PCR Real-Time PCR Gene cyclers available from many suppliers
28.
©2001 Timothy G.
Standish 05/30/15 28 Multiplex PCRMultiplex PCR PCR reactions can be devised in which several targets are amplified simultaneously — often used in diagnostic applications.
29.
©2001 Timothy G.
Standish 05/30/15 29 What is RT-PCRWhat is RT-PCR RT-PCR is Reverse transcription PCR The source of material is mRNA First step is cDNA synthesis by reverse transcriptase at 42ºC Second step is Standard PCR procedure Result is cDNA of gene target Reverse-trascriptionPCRReverse-trascriptionPCR
30.
©2001 Timothy G.
Standish 05/30/15 30 PCRPCR 5’ 3’ 5’3’ 5’ 3’ 5’3’ 5’ 3’ 5’3’ 5’ 3’ 5’3’ 5’ 3’ 5’3’ 5’ 5’ 3’ 3’ 5’ 3’ 5’3’ 5’ 3’5’3’ denaturation (94 o C) primer annealing (50-70 o C) primer extension (72 o C) Next round….. 3’ RT-PCRRT-PCR 5’ AAAAAn TTTTTn 5’ 3AAAAAn TTTTTn first-strand cDNA synthesis by RT 53’ TTTTTn 53’ 5’ TTTTTn 5’ AAAAAn RNase treatment; primer annealing (50-70 o C) primer extension (72 o C) mRNA TTTTTn3’ 5’ 5’ AAAAAn TTTTTn AAAAAn Next round….. An example was described in the article by Halpin et al. (1998). Reverse-trascriptionPCRReverse-trascriptionPCR Basic Reaction of PCR & RT-PCRBasic Reaction of PCR & RT-PCR
31.
©2001 Timothy G.
Standish 05/30/15 31 Applications of PCRApplications of PCR Mutation testing, e.g. cystic fibrosis. Diagnosis or screening of acquired diseases, e.g. AIDS. Genetic profiling in forensic, legal and bio- diversity applications. Site-directed mutagenesis of genes. Quantitation of mRNA in cells or tissues.
32.
THE END THANKS FORYOUR ATTENTION
Jetzt herunterladen