genetics recombinant dna amp genomic technology

... steps used to clone DNA?     Isolate DNA from an organism Cut the organismal DNA and the vector with restriction enzymes making recombinant DNA Introduce the recombinant DNA into a host Restriction ... of DNA Using DNA probes to screen plasmid genomic libraries for specific DNA sequences PowerPoint® Layered Art Figure 8.11 Using DNA probes to screen plasmid genomic libraries for specific DNA ... to produce DNA fragments of appropriate size for constructing a genomic library cDNA Libraries are best for eukaryotes Figure 8.8 The synthesis of double-stranded complementary DNA (cDNA) from...

Ngày tải lên: 13/03/2014, 19:33

... using recombinant DNA technology was human insulin, developed by Genentech and Licensed by Eli Lilly and Company  Applications of recombinant biotechnology  Recombinant human insulin  Recombinant ... successful production and intracellular replication of recombinant DNA appeared in 1972 and 1973 Stanford University applied for a US patent on recombinant DNA in 1974, listing the inventors as Stanley ... Self-replicating  Double stranded  Circular DNA  Independent extrachromosomal DNA in  Ampicillin bacteria 1kb-500kb  Kanamycin  Origin of DNA replication, ori  High (10-100) & low copy...

Ngày tải lên: 23/03/2014, 22:20

... DNA pieces are joined in vitro to form recombinant molecules • Generate sticky ends on the DNA, e.g with restriction endonucleases • Tie DNA molecules from different sources together with DNA ... CTTAA G Recombinant molecules G AATTC CTTAA G GAATTC CTTAAG GAATTC CTTAAG Parental molecules DNA ligase covalently joins two DNA molecules • Uses ATP or NADH to provide energy to seal nicks DNA ligase ... cycles, the amplification is approximately [2exp(n-1)]-2 • After 21 cycles, the fragment has been amplified about a million-fold • E.g a sample with 0.1 pg of the target fragment can be amplified...

Ngày tải lên: 13/03/2014, 18:39

... host cell population have no recombinant DNA replicating in them This selection or screening for desired recombinants provides a route to recovery of the recombinant DNA of interest in pure form ... cloning of yeast DNA, or certain prokaryotic DNAs, in yeast A vehicle for recombinant DNA and a transformation procedure are also available for Staphylococcus aureus.S2 Many recombinants containing ... a Tc s recombinant plasmids survive 29"~3 Similarly, the PstI site of pBR322 is within the Ap gene Insertion of DNA there creates an Ap s Tc R plasmid, a3 However, most rat insulin cDNA recombinant...

Ngày tải lên: 11/04/2014, 10:25

... degree of purity of the DNA substrate Improperly prepared DNA samples will be cleaved poorly, if at all, producing partially digested DNA In addition to DNA purity, other DNA- associated parameters ... plasmid or viral DNAs completely than for linear DNA A comparison of the relative cleavage efficiencies for several supercoiled and linear DNAs are presented in Table V If a supercoiled DNA (e.g., ... lambda DNA under standard reaction conditions ' All enzymes and DNAs were from Bethesda Research Laboratories, Inc d Linear form III pBR322 DNA was prepared by incubation of supercoiled form I DNA...

Ngày tải lên: 11/04/2014, 10:25

... of nematode DNA fragments produce recombinant phages that are detected by plating on Q359 The yield of recombinants is linear with the amount of nematode DNA added as long as the DNA concentration ... of insert DNA to vector DNA (0.5/~g insert DNA per 1.0/~g of vector) The yield of recombinants in this experiment ranged from 2.4 x 10~ to 5.4 x 105 per microgram of 15-20 kb nematode DNA This ... suggestion by one of us DNA is cotransformed into mammalian cells together with su+~H DNA to provide a selective marker Larger DNA fragments carrying transforming DNA and the suCH DNA are then selected...

Ngày tải lên: 11/04/2014, 10:26

... of ssDNA Selective Packaging of the (Recombinant) DNA Strands of pKUN The production of ss recombinant DNA with this system in many respects resembles that for obtaining recombinant M13 DNA 1°,12Therefore ... prokaryotic origin Some examples of packaging of ss recombinant plasmid DNA are shown in Fig 3A The level and ratio of ss phage DNA (upper band in Fig 3A) to ss plasmid DNA (lower band) vary for ... ss phage DNA and the other ss plasmid DNA (see Fig 3A) Sometimes you might also see traces of high-molecular-weight DNA In case of packaging by IKe-9 another, fast migrating, (miniphage DNA) band...

Ngày tải lên: 11/04/2014, 10:26

... RNA strand is converted to DNA by nicktranslation repair catalyzed by Escherichia coli DNA polymerase I, RNase H, and DNA ligase The end product, a closed circular cDNA recombinant, is transfected ... OF ~ DNA POLYMERASE1 AND DNA UGASE J /H~m DIGESTION Xho[8BamHl GGc~'~-~ "~" Hk'~dH] 'P~ tl~ A TAA FIG Enzymatic steps in the construction of pcD-cDNA recombinants The designations of the DNA segments ... high molecular weight DNA by base-pairing between dG-tails, the yield of linker DNA from agarose gels is generally poor cDNA Cloning Steps in the construction of cDNA-plasmid recombinants are illustrated...

Ngày tải lên: 11/04/2014, 10:26

... Site-specific DNA methylases (S-adenosylmethionine : DNA methyltransferases) have a variety of uses in DNA analytical procedures Methylases may be used as probes for DNA conformation,L2 in recombinant DNA ... sequence specificities Materials Restriction endonucleases, purified DNA methylases, T4 DNA ligase, unmethylated phage h DNA, ~bX174 RF I DNA, and 5'-phosphorylated oligonucleotides were obtained from ... and selective DNA methylase assay early in many DNA methylase purifications For example, the M.MbolI methylase must be assayed in the presence of M.MboI and at least three other DNA methylases...

Ngày tải lên: 11/04/2014, 10:26

... of DNA are captured from large sample volumes, e.g., 10 ng of DNA from 0.5 ml of sample; (5) relatively large amounts of DNA are captured from small volumes, e.g., /~g of DNA from 30 /zl of sample; ... lentil; lane 6, soybean DNAs and were prepared from crushed green leaves; DNA from flour; DNAs 4-6 from crushed seeds The electrophoresis run conditions were as follow: time ramp 50 to 200 sec at ... allowed for the digestion of DNA in solution Microbeads not only protect large DNA molecules from shear, but also act as giant DNA- carrying "cells," thus converting DNA into a "solid state" and...

Ngày tải lên: 11/04/2014, 10:26

... Cloning of DNA Fragments Any of several reliable methods can be used to prepare DNA/ cDNA for cloning in pIFFS For direct in-frame cloning in pIFF8 (prepared as above) the foreign DNA fragment ... not will remain Lac + Examples Selection of Open Reading Frames in DNA/ cDNA The pIFF series of vectors, and in particular pIFF8, are recent vector constructions and examples on experimental applications ... expression [4] M D R I GENE IN MAMMALIAN EXPRESSION VECTORS 45 DNA Detection Transfected MDR1 DNA can be detected by isolating genomic DNA from multidrug-resistant cells and performing Southern analysis...

Ngày tải lên: 11/04/2014, 10:26

... (1991) FIG Sample sequencing ladders obtained from PCR-amplified ssDNA (A-C) Amplifications from Mus spretus genomic DNA; (D-F) amplifications from Drosophila melanogaster genomic DNA (A and B) ... product directly from genomic DNA relies on having an efficient method for sequencing the amplified DNA Several approaches have been taken for sequencing PCR-amplified DNA The ease and reproducibility ... Acad Sci U.S.A 86, 6196 (1989) a 91011 121314 dsDNA b 91011121314 dsDNA ssDNA =~ C dsDNA =~ 91011121314 [1] SEQUENCING OF in Vitro AMPLIFIED DNA 50/1 50/2 50/3 50/4 5o/5 1150 2/50 3/50 4•50...

Ngày tải lên: 11/04/2014, 10:27

... conclusion, new technology such as GWAS has the potential to improve our understanding of genetics of sepsis Large studies will have to be conducted, however, to effectively utilize this technology ... the role of genetic variants in the spectrum of infection and severe sepsis susceptibility For example, comparing the distribution of the genetic variant in severe sepsis cases and healthy, population-based...

Ngày tải lên: 13/08/2014, 15:21

... 50 Activity 7.2: Determining the Impact of Technology on a Windows DNA Design Exercise 1: Determining Technology Implications ! Determine technology implications Participate in small groups ... instructor Identify the level of impact — high, medium, or low — that each technology type would have on the given layer of a Windows DNA design Write your answers in the grid provided After completing ... chart Use the space below for brainstorming Activity 7.2: Determining the Impact of Technology on a Windows DNA Design 51 Write your answers in the table below User Interface User Services Business...

Ngày tải lên: 21/12/2013, 06:16

... the CRPW13F(cAMP)2 complex, the concentration of cAMP was 200 lM, whereas in the case of CRPW13F(cAMP)4 the concentration of cAMP was mM The molar ratio CRP to DNA in the proteinDNA complex was ... Trp13 microenvironments on cAMP binding to anti-cAMP sites may play a signicant role in the CRPCytRDNA complex In contrast with Trp13 of CRP, binding of cAMP to anticAMP-binding sites does not ... cAMP concentration, there are two identical interactive high-afnity sites for cAMP and at least one low-afnity cAMP-binding site at high concentration of the ligand [8] The idea of the four cAMP-binding...

Ngày tải lên: 20/02/2014, 23:20

... orders@intechopen.com DNA Methylation – From Genomics to Technology, Edited by Tatiana Tatarinova and Owain Kerton p cm ISBN 978-953-51-0320-2 Contents Preface IX Part Epigenetics Technology and Bioinformatics ... 2008, Current Genomics, Vol 9, pp 394-408 X Preface and the control of tissue-specific gene expression DNA methylation also has health implications, for example the gain or loss of DNA methylation ... Background DNA Methylation was initially addressed as one of the most primitive mechanisms that organisms utilize to (a) protect genomic DNA and initiate the host resistance mechanism towards foreign DNA...

Ngày tải lên: 08/03/2014, 19:20

... e¢¢) and then exposed to DNase I Whole genomic DNA (lane a) and DNase I-digested genomic DNA (lane b) were controls (B) Electrophoretic migration, at 37 °C, of genomic DNA preincubated with 600 ... I Controls were the whole genomic DNA (lane a) and the DNA fully digested by DNase I (lane b) The DNA kb ladder marker was in lane m Fig Electrophoresis of genomic DNA preincubated with single ... naked genomic DNA from DNase I To assess the protective role of NAPs against endonucleases and compare it with that of single polyamines, we carried out electrophoretic assays of genomic DNA treated...

Ngày tải lên: 23/03/2014, 15:20

... reverse transcribed into its DNA complement (complementary DNA, or cDNA) using the enzyme reverse transcriptase mRNA Reverse transcriptase cDNA Taq DNA polymerase DNA  Oligo dT(18) or Random ... 4oC, 0s  Exponentially amplifying a single or some piece of DNA generating thousands of million copies of DNA  Total copies = 2n where n is the number of cycles to copy DNA  So basically it ... PCR reaction  Template DNA  Primers  dNTPs  Taq DNA polymerase  Buffer  Steps programmed in PCR  Step 1: Denature DNA, 95oC, 5min  Step 2: Annealing: - Denature DNA, 950C, 30s-1min - Annealing,...

Ngày tải lên: 23/03/2014, 22:20

... Primrose, S B & Twyman, R M (2006a) Applications of recombinant DNA technology In Primrose, S B & Twyman, R M (Eds.) Principles of gene manipulation and genomics Oxford, Blackwell Publishing Primrose, ... production of many human recombinant proteins, for example blood clotting factors, cytokines, growth factors, immunoglobulins, and thrombolytics (Chun et al., 2001) ANIMAL CELL TECHNOLOGY Animal cell ... foreign DNA into animal cells  Direct transfer by physical transfection Using physical transfection methods, such as microinjection, particle bombardment, ultrasound, and electroporaion, naked DNA...

Ngày tải lên: 28/03/2014, 09:20

... Gina The genomics age : how DNA technology is transforming the way we live and who we are / Gina Smith p cm Includes index ISBN 0-8144-0843-5 Genetics Popular works Genomics—Popular works Genetics Social ... discoveries scientists have made about DNA could change your life, your health, and society The DNA Files The most important advance to come out of his work, says DNA double helix discoverer James ... determining how the DNA in every human being affects identity, health, and disease And genomics is launching other sciences almost as quickly as you can learn the terms First functional genomics, then...

Ngày tải lên: 08/04/2014, 13:10