A mathematical model and a computerized simulation of PCR using complex templates Nucleic Acid Research, 1996, Vol. 24, No. 18, pp. 3538-3545 E. Rubin.

Slides:



Advertisements
Similar presentations
RAPD Randomly Amplified Polymorphic DNA
Advertisements

Nucleic Acid Hybridization Nucleic acid hybridization is a fundamental tool in molecular genetics which takes advantage of the ability of individual single-stranded.
Tools for Molecular Biology Amplification. The PCR reaction is a way to quickly drive the exponential amplification of a small piece of DNA. PCR is a.
Genome Scale PCR Infidelity Search Goal: An efficient search for the presence of potential undesired PCR products that scans through 3 billion bases of.
PCR - Polymerase Chain Reaction PCR is an in vitro technique for the amplification of a region of DNA which lies between two regions of known sequence.
Python Programming on PCR Primers Design
PCR Polymerase Chain Reaction. PCR - a method for amplifying (copying) small amount of DNA in nearly any amount required, starting with a small initial.
Basics of hybridization. What is hybridization? n Complementary base pairing of two single strands of nucleic acid  double strand product u DNA/DNA u.
Real-Time Primer Design for DNA Chips Annie Hui CMSC 838 Presentation.
Center for Homogeneous DNA Analysis - Software April 21, 2005.
Nucleic Acid Design Applications Polymerase Chain Reaction (PCR) Calculating Melting Temperature (Tm) PCR Primers Design.
Sequence Analysis. DNA and Protein sequences are biological information that are well suited for computer analysis Fundamental Axiom: homologous sequences.
©2003/04 Alessandro Bogliolo Primer design. ©2003/04 Alessandro Bogliolo Outline 1.Polymerase Chain Reaction 2.Primer design.
Interdisciplinary Center for Biotechnology Research
Accuracy: The closeness of a measured volume to the true volume as specified by the volume setting of the pipette. Also known as “mean error”. precision:
Variants of PCR Lecture 4
PCR Primer Design Guidelines
IN THE NAME OF GOD. PCR Primer Design Lecturer: Dr. Farkhondeh Poursina.
PCR optimization. Primers – design must be good but influenced by template sequence Quality of template DNA/impurities Components of PCR may need to be.
Nucleic Acid Secondarily Structure AND Primer Selection Bioinformatics
The methods used by molecular biologists to study DNA have been developed through adaptation of the chemical reactions and biological processes that occur.
PCR Modeling MEC Lim Hee Woong.
DIY Primer Design Oligonucleotides for Special Applications in Molecular Biology Alberto Catalano Kanematsu Labs, Institute of Haematology RPAH
PCR By Staci Cutting and Mitch Gavazzi. What is PCR? PCR is sometimes called Molecular photocopying the polymerase chain reaction is a fast and inexpensive.
Tools of Bioinformatics
Polymerase Chain Reaction PCR. invented by Karry B. Mullis (1983, Nobel Prize 1993) patent sold by Cetus corp. to La Roche for $300 million depends on.
Dave Palmer Primer Design Dave Palmer
Primer Design and Computer Program Does it really matter? Principles of Primer Design Can I trust my gut feeling? What should I do? Sean Tsai ©1999, National.
Amplification of Genomic DNA Fragments OrR. Amplification To get particular DNA in large amount Fragment size shouldn’t be too long The nucleotide sequence.
Polymerase Chain Reaction (PCR) Developed in 1983 by Kary Mullis Major breakthrough in Molecular Biology Allows for the amplification of specific DNA fragments.
Design of oligonucleotides for microarrays and perspectives for design of multi-transcriptome arrays Henrik Bjorn Nielsen, Rasmus Wernersson and Steen.
© 2004, SNU Biointelligence Lab, Joining and Rotating Data with Molecules Masanori Arita, Masami Hagiya and Akira Suyama Summarized.
CHAPTER SIX Nucleic acid hybridization: principles and applications 생물정보학협동과정 강민호.
Denaturation temperatuare gradient- polymerase chain reaction: by-product? Ji Youn Lee Cell and microbial engineering laboratory Seoul National University.
Human Genomics. Writing in RED indicates the SQA outcomes. Writing in BLACK explains these outcomes in depth.
Computational and experimental analysis of DNA shuffling : Supporting text N. Maheshri and D. V. Schaffer PNAS, vol. 100, no. 6, Summarized by.
The Inference via DNA Computing Piort Wasiewicz et al. Proceedings of the 1999 Congress on Evolutionary Computation, vol. 2, pp Cho, Dong-Yeon.
A Software Tool for Generating Non-Crosshybridizing libraries of DNA Oligonucleotides Russell Deaton, junghuei Chen, hong Bi, and John A. Rose Summerized.
BioIntelligence Lab Reaction discovery enabled by DNA-templated synthesis and in vitro selection Matthew W. Kanan, Mary M. Rozenman, Kaori Sakurai, Thomas.
Probe Selection Problems in Gene Sequences. (C) 2003, SNU Biointelligence Lab, DNA Microarrays cDNA: PCR from.
From: Duggan et.al. Nature Genetics 21:10-14, 1999 Microarray-Based Assays (The Basics) Each feature or “spot” represents a specific expressed gene (mRNA).
Polymerase Chain Reaction: A Markov Process Approach Mikhail V. Velikanov et al. J. theor. Biol Summarized by 임희웅
Basics of hybridization. What is hybridization? n Complementary base pairing of two single strands of nucleic acid  double strand product u DNA/DNA u.
D. Darban, Ph.D Department of Microbiology School of Medicine Alborz University of Medical Sciences 1 Probe and Primer Design.
Molecular Evolutionary Computing (MEC) for Maximum Clique Problems March 9, 2004 Biointelligence Laboratory School of Computer Science and Engineering.
1 Summarized by Ji Youn Lee. 2 Model Development.
Fac. of Agriculture, Assiut Univ.
Polymerase Chain Reaction
Gel electrophoresis analysis Automated DNA analyzer.
PCR TECHNIQUE
Polymerase Chain Reaction & DNA Profiling
Selection of Oligonucleotide Probes for Protein Coding Sequences
Lecture 4: Probe & primer design
Sequences and their Properties
PNA-mediated Whiplash PCR
Polymerase Chain Reaction
B3- Olympic High School Bioinformatics
Nucleic Acid Interactions Practicalities
Fuzzy logic with biomolecules
Introduction to Bioinformatics II
Summarized by In-Hee Lee
Oligonucleotide sequences of polymerase chain reaction (PCR) primers and competitive templates. Oligonucleotide sequences of polymerase chain reaction.
Fitness measures for DNA Computing
Information Theoretical Probe Selection for Hybridisation Experiments
Maximal Clique Problem with Experiment
Sequences and their Properties
Russell Deaton, junghuei Chen, hong Bi, and John A. Rose
Nearest-neighbor model for Tm
Genomic structure of LTBP-4 around the 3rd 8-Cys repeat.
Designs for Autonomous Unidirectional Walking DNA Devices
Presentation transcript:

A mathematical model and a computerized simulation of PCR using complex templates Nucleic Acid Research, 1996, Vol. 24, No. 18, pp E. Rubin and A. A. Levy Summarized by Shin, Soo-Yong

(C) 2001, SNU Biointelligence Lab, Bric biosoftware homepage db/biosoft/index_1.html

(C) 2001, SNU Biointelligence Lab, PCR primer check program Primer premier  PCR 을 위한 primer 및 oligonucleotide probe 를 선택 및 분석하 는 프로그램.  PrimerCheck PCR 을 위한 primer 및 oligonucleotide probe 의 물리적인 특성을 알려 주는 프로그램. 용도 및 특징을 정리하면 다음 과 같다. A, C, G and T contents absolute and CG contents in Calculates annealing temperature Calculates molweight of the primer. 

(C) 2001, SNU Biointelligence Lab, PCR primer check program PrimerDesign PCR 을 위한 primer 및 oligonucleotide probe 를 선택 및 분석하는 dos 용 프로그램. 용도 및 특징을 정리하면 다음과 같다. 1) 새로운 primer 쌍을 합성시 사용 2) 한 방향의 주어진 primer 를 가지고 다른 방향의 primer 를 합성시 사용 3) 한 염기 서열 중 반복되는 서열을 찾고자 할 때 사용 4) 한 염기 서열 안의 고유한 서열을 찾고자 할 때 사용 5) 32,000 bp 정도의 염기 서열도 이용가능 

(C) 2001, SNU Biointelligence Lab, PCR simulation Amplify 2.5  알고 있는 DNA 서열에 대하여 선택하거나 새로 입력한 primer 로 PCR 을 수행하였을 때의 결과를 예측해주는 프로그램이다. PCR 결과로 만들어 지 는 band 의 크기를 예상하고 primer 가 template 에 얼마나 잘 annealing 하는지 예측해 준다.  y/index.html  문제점 : MacOS

(C) 2001, SNU Biointelligence Lab, Annhyb Oligonucleotide 서열을 5′ 에서 3′ 방향으로 적어주면, nearest neightboors formula 에 의해 oligonucleotide 의 Tm 값을 구해 준다. Salt 나 primer 의 양을 변화 시키면 변 화된 값에 대한 Tm 값이 자동적으로 계산되어 나타난다. 결 과는 file 형태로 저장할 수 있다.Degenerated 서열에 대해 서는 Tm determination 이라는 버튼이 자동적으로 생기며 이를 click 하면, 구분된 서열에 대한 Tm 의 최소값과 최대값 을 구할 수 있다. Revert complement 서열을 얻고자 하는 서열을 5′ 에서 3′ 방향으로 입력한 뒤, revert- complement 버튼을 눌러서 원하는 결과를 얻는다. 

(C) 2001, SNU Biointelligence Lab, References in paper BIND  Hartemink et al. Oligo  HYBsimulator   Bronken link

(C) 2001, SNU Biointelligence Lab, References in paper HYTHER   서버 다운 PCRsim  발표할 paper Amplify  Engels, W. R. (1993) Trens in Biochemical Sciences 18,

(C) 2001, SNU Biointelligence Lab, Primer genome.wi.mit.edu/ftp/distribution/software/primer.0.5/ genome.wi.mit.edu/ftp/distribution/software/primer.0.5/ PRIMER is a computer program for automatically selecting PCR primers. PRIMER tests oligos for annealing temperature, complementarity to genomic repeat sequences (e.g. Alu), ability to form primer-dimer, and other criteria.

(C) 2001, SNU Biointelligence Lab, References in paper Reif  Not available Hagiya et al.  SCAN  Hatermink

(C) 2001, SNU Biointelligence Lab, Tm 계산 site meltinghome.html meltinghome.html Melting homepage

(C) 2001, SNU Biointelligence Lab, The expected probability of obtaining a non-targeted PCR product under stringent annealing conditions is extremely low The frequent amplification of non-targeted products in real PCR is not caused by deviations from randomness in nucleotides order or composition, but rather by the tolerance of PCR to mismatches Mismatch tolerance is the most significant factor affecting PCR specificity, followed by primer length, template size and product size limit

(C) 2001, SNU Biointelligence Lab, mPCR.html mPCR.html Broken link

(C) 2001, SNU Biointelligence Lab, A model for PCR Conditions  The template made of 4 nt in an equal ratio and a random order  Annealing may occur at any site of the template which is similar to the primer within mismatch tolerance  Any two opposing sites within product size limit give a PCR product

(C) 2001, SNU Biointelligence Lab, A model for PCR N pairs : total number of annealing site pairs XY-products : 2N pairs X or Y-products : N pairs What is end effect?

(C) 2001, SNU Biointelligence Lab, PCR probability The probability that a site will anneal to a specific primer with precisely m mismatches p(l,m)  Binomial distribution  l experiments, l-m success, m fail

(C) 2001, SNU Biointelligence Lab, The number of PCR products

(C) 2001, SNU Biointelligence Lab, The number of PCR products

(C) 2001, SNU Biointelligence Lab, The number of annealing sites for both primers

(C) 2001, SNU Biointelligence Lab, PCR with real primers DB search? What does it mean? Virtual PCR  M. Lexa, J. Horak, and B. Brzobohaty, Bioinformatics, Vol. 17, no. 2, pp

(C) 2001, SNU Biointelligence Lab, 확인하고 싶은 논문 Tae-Ju Cho and Sang-Soo Park, A simulation of subtractive hybridization, Nucleic Acis Research, 1998, Vol. 26, No. 6, pp