pharmacology recombinant dna technology—biotechnology

... 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 ... Can carry up to 25kbDNA  1000 time more efficient than a plasmid Cloning in Lambda Vectors Make a partial digest of DNA Ligate the DNA to the arms of the vector Package the DNA into phage particles...

Ngày tải lên: 23/03/2014, 22:20

... 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 ... RNase H, DNA polymerase I, and DNA ligase Cloning of cDNA using BamHI linkers Screening     What screening method can be used to identify a recombinant clone with a target gene in a cDNA library ... 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...

Ngày tải lên: 13/03/2014, 19:33

... 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 ... 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, ... Restriction digestion of genomic DNA Agarose gel electrophoresis Denature DNA Transfer ssDNA to membrane Membrane baking Hybridization Wash and detection  DNA denature Denaturation is a process...

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 ... nick T4 DNA ligase + ATP P P P P P P P P P A G G A A T T C T C C T T A A G P P P P P P G T P P P A A C T P P Alternate method to join DNA: homopolymer tails Alternate method to join DNA: linkers...

Ngày tải lên: 13/03/2014, 18:39

... 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 ... atom or going to the moon.”1 Introducing Your DNA As most everyone knows by now, DNA is short for deoxyribonucleic acid But you know where your DNA is? Can you tell a gene from a chromosome? ... companies are using worm DNA to figure out how to slow human aging, or how doctors are employing DNA knowledge to finally win the fight against cancer? Most people don’t The DNA sciences will dominate...

Ngày tải lên: 08/04/2014, 13:10

... 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 ... 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 ... plasmid DNA was observed although phage DNA was produced To ensure optimal production of ssDNA we recommend initially trying infections with several helper phages Even if the amount of plasmid DNA...

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 ... with 1/zg of ~bX174 RF I DNA in the standard restriction endonuclease assay buffer in a volume of 0.05 ml The supercoiled ~bX174 DNA has been substituted for Adeno-2 DNA because neither NotI...

Ngày tải lên: 11/04/2014, 10:26

... molecular weight DNA molecules, socalled megabase DNA (Mbp DNA) For various reasons, plant molecular biologists have been slow to develop specific protocols suitable for preparing Mbp DNA Without ... 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 ... nucleic acid from the DNA capture reagent Thus, we have analyzed the released DNA using procedures that not require native DNA Results Basic Features of Capture and Release of DNA We initially performed...

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 ... Vector and DNA Fragments with Sticky Ends Mix 100 ng of prepared vector (Section l) and target DNA in a molar 18 VECTORS FOR EXPRESSING CLONED GENES [2] ratio of : with I U of T4 DNA ligase (1 ... Procedure 2.2: Ligation of Vector and DNA Fragments with Blunt Ends Mix 200 ng of prepared vector (Section 1) and target DNA in a molar ratio of : with 12 U of T4 DNA ligase (8 U//A) (efficient blunt-end...

Ngày tải lên: 11/04/2014, 10:26

... Lee, DNA 10, 67 (1991) 6 METHODSFOR SEQUENCINGDNA [1] Single-Stranded DNA Templates Sequencing problems derived from strand reassociation can be avoided by preparing single-stranded DNA (ssDNA) ... 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 ... exhausted, ssDNA is produced The ssDNA produced can be sequenced either using the limiting PCR primer or an internal primer complementary to the ssDNA reaction products on a gel The ssDNA yield...

Ngày tải lên: 11/04/2014, 10:27

Ngày tải lên: 04/06/2014, 15:17

... antibodies 18 1.4.1 Lipids as disease biomarkers 18 1.4.2 Recombinant phage display 20 1.4.3 Recombinant antibody expression 21 ii Table of Contents 1.5 Lipid ... 4',6-diamidino-2-phenylindole (DNA binding fluorescent dye) DC dendritic cell DIC differential interference contrast DMPA dimyristoyl phosphatidic acid DMPC dimyristoyl phosphocholine DNA deoxyribonucleic ... acid fast staining (5,36) In particular, line probe assays, in which labelled amplified DNA is hybridized to DNA probes along a strip membrane, are fast and highly accurate (57,61) This has led...

Ngày tải lên: 10/09/2015, 09:05

... 50 Activity 7.2: Determining the Impact of Technology on a Windows DNA Design Exercise 1: Determining Technology Implications ! Determine technology implications Participate ... 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 the above steps, you will discuss ... 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 Services...

Ngày tải lên: 21/12/2013, 06:16

... hidden picture ModellingMethylation Dynamics Modelling DNA DNA Methylation Dynamics 2.1.1 Methods to analyse DNA patterns Since the advent of DNA sequencing technologies, (França et al., 2002), ... protein -DNA interactions combining chromatin immuno precipitation and massively parallel DNA sequencing techniques or microarray (chip) experiments ModellingMethylation Dynamics Modelling DNA DNA ... loci of “foreign” DNA insertions, which tend to disturb the existing DNA methylation patterns, (Collins et al., 1998) Information from literature indicates that a majority of DNA methylation occurs...

Ngày tải lên: 08/03/2014, 19:20

... TECHNOLOGY -VOLUME Advanced scientific computing in BASIC with applications in chemistry, biology and pharmacology DATA HANDLING IN SCIENCE AND TECHNOLOGY Advisory Editors: B.G.M Vandeginste, O.M Kvalheim ... Morgan Volume Advanced Scientific Computing in BASIC with Applications in Chemistry, Biology and Pharmacology by P Valk6 and S.Vajda DATA HANDLING IN SCIENCE AND TECHNOLOGY -VOLUME Advisory Editors: ... and L Kaufman Advanced scientific computing in BASIC with applications in chemistry, biology and pharmacology P VALKO Eotvos Lorand University, Budapest, Hungary S VAJDA Mount Sinai School of Medicine,...

Ngày tải lên: 22/03/2014, 23:20

Ngày tải lên: 24/03/2014, 04:20