For the Tutorial Programme in Proteomics High
... Restriction enzymes type II and ligases. These two sets of enzymes have complementary activity, restriction enzymes work as “scissors” capable of identifying and cleaving specific DNA sequences (Kelly and Smith 1970, Smith and Wilcox 1970), whereas ligases re-join two DNA strands and reconstitute th ...
... Restriction enzymes type II and ligases. These two sets of enzymes have complementary activity, restriction enzymes work as “scissors” capable of identifying and cleaving specific DNA sequences (Kelly and Smith 1970, Smith and Wilcox 1970), whereas ligases re-join two DNA strands and reconstitute th ...
GENETICS
... how the organization contributes to both continuity and variability in the genetic information. Explain some mechanisms by which gene expression is regulated in prokaryotic and eukaryotic genomes. Explain current recombinant technologies. Explain some practical applications of nucleic acid technolog ...
... how the organization contributes to both continuity and variability in the genetic information. Explain some mechanisms by which gene expression is regulated in prokaryotic and eukaryotic genomes. Explain current recombinant technologies. Explain some practical applications of nucleic acid technolog ...
recombinant dna technology and genetic engineering
... sources with the same restriction endonuclease (BamHI in this case). BamHI cuts the same site on both molecules 5' GGATCC 3' 3' CCTAGG 5'. The ends of the cut have an overhanging piece of single-stranded DNA. These are called "sticky ends" because they are able to base pair with any DNA molecule con ...
... sources with the same restriction endonuclease (BamHI in this case). BamHI cuts the same site on both molecules 5' GGATCC 3' 3' CCTAGG 5'. The ends of the cut have an overhanging piece of single-stranded DNA. These are called "sticky ends" because they are able to base pair with any DNA molecule con ...
DNA Technology20082009
... – a set of techniques for combining genes from different sources into a single DNA molecule producing recombinant DNA. • An organism that carries recombinant DNA is called a genetically modified (GM) organism. ...
... – a set of techniques for combining genes from different sources into a single DNA molecule producing recombinant DNA. • An organism that carries recombinant DNA is called a genetically modified (GM) organism. ...
2016 Midterm answer key
... experiment that distinguishes between these two possibilities. That is, design an experiment which will determine whether or not the ability of any individual colony recovered in your selection not only has a plasmid but that growth at 37C is dependent upon the presence of that specific plasmid. To ...
... experiment that distinguishes between these two possibilities. That is, design an experiment which will determine whether or not the ability of any individual colony recovered in your selection not only has a plasmid but that growth at 37C is dependent upon the presence of that specific plasmid. To ...
High efficiency of site-directed mutagenesis mediated by a single
... annealed to the denatured plasmid and directs the synthesis of the mutant strand. After digestion with selection enzyme, the plasmid DNA is amplified into Escherichia coli strain BMH71-18 and subjected to a second digestion and amplification into the bacterial strain DH5α. A mutagenesis efficiency > ...
... annealed to the denatured plasmid and directs the synthesis of the mutant strand. After digestion with selection enzyme, the plasmid DNA is amplified into Escherichia coli strain BMH71-18 and subjected to a second digestion and amplification into the bacterial strain DH5α. A mutagenesis efficiency > ...
Activity #5b. Plasmid DNA Isolation, Restriction Enzyme Digestion
... C2. Decant (pour off) the supernatant into your liquid waste container. (Ensure that RNase A has been added to Buffer P1—Your Professor or TA will probably have done this for you already) Resuspend both pellets of bacterial cells in a total of 250 µl Buffer P1: --put the 250 uL Buffer P1 into one tu ...
... C2. Decant (pour off) the supernatant into your liquid waste container. (Ensure that RNase A has been added to Buffer P1—Your Professor or TA will probably have done this for you already) Resuspend both pellets of bacterial cells in a total of 250 µl Buffer P1: --put the 250 uL Buffer P1 into one tu ...
Molecular and General Genetics
... plification during the starvation phase (Fig. la). Earlier experiments showed that Bacillus fl-glucanase was mainly accumulated within the periplasmic space of E. coli cells (Cantwell and McConnell 1983; Borriss et al. 1985). We found that the overexpression of otherwise periplasmic flglucanase in E ...
... plification during the starvation phase (Fig. la). Earlier experiments showed that Bacillus fl-glucanase was mainly accumulated within the periplasmic space of E. coli cells (Cantwell and McConnell 1983; Borriss et al. 1985). We found that the overexpression of otherwise periplasmic flglucanase in E ...
Making Recombinant DNA
... DNA species makes the circular plasmid DNA denser than the chromosomal DNA, the plasmids form a distinct band on centrifugation in a cesium chloride gradient and can be separated easily. They can then be introduced into bacterial cells by transformation. Restriction enzymes: Have two properties usef ...
... DNA species makes the circular plasmid DNA denser than the chromosomal DNA, the plasmids form a distinct band on centrifugation in a cesium chloride gradient and can be separated easily. They can then be introduced into bacterial cells by transformation. Restriction enzymes: Have two properties usef ...
In vitro formation of a catabolic plasmid carrying
... was isolated from a selection of 12 such colonies and was analysed by digestion with HindIII. Each isolate carried a 9 kb HindIII insert. Digestion with EcoRI gave either two approximately 7 kb bands (four colonies) or an 11 kb band plus a 3 kb band (eight colonies). These patterns suggested that th ...
... was isolated from a selection of 12 such colonies and was analysed by digestion with HindIII. Each isolate carried a 9 kb HindIII insert. Digestion with EcoRI gave either two approximately 7 kb bands (four colonies) or an 11 kb band plus a 3 kb band (eight colonies). These patterns suggested that th ...
Chapter 6
... or genetic engineering or—Cohen’s choice—genetic manipulation. “Genetic manipulation,” he wrote in Scientific American, “opens the prospect of constructing bacterial cells which can be grown easily and inexpensively, that will synthesize a variety of biologically produced substances such as antibiot ...
... or genetic engineering or—Cohen’s choice—genetic manipulation. “Genetic manipulation,” he wrote in Scientific American, “opens the prospect of constructing bacterial cells which can be grown easily and inexpensively, that will synthesize a variety of biologically produced substances such as antibiot ...
Chapter 20 Practice Multiple Choice
... of a thymine kinase receptor. A new drug (Gleevec or imatinib) has been found to inhibit the disease if the patient is treated early. ____ 22. Why would Gleevec most probably cause remission of the disease? a. It reverses the chromosomal translocation. b. It eliminates the Ph' chromosome. c. It remo ...
... of a thymine kinase receptor. A new drug (Gleevec or imatinib) has been found to inhibit the disease if the patient is treated early. ____ 22. Why would Gleevec most probably cause remission of the disease? a. It reverses the chromosomal translocation. b. It eliminates the Ph' chromosome. c. It remo ...
Mammalian Two-Hybrid Assay Kit
... for protein-protein interactions in various mammalian cell lines. The cells must not contain molecules or proteins that interfere with the interactions between the proteins of interest, and must contain proteins or enzymes that may be necessary for the interactions. Background expression levels of t ...
... for protein-protein interactions in various mammalian cell lines. The cells must not contain molecules or proteins that interfere with the interactions between the proteins of interest, and must contain proteins or enzymes that may be necessary for the interactions. Background expression levels of t ...
SBI 3U Genetics Review Questions LG #1: DNA
... 2. Identify the structures of adenine, guanine, thymine, and cytosine. Which nucleotides pair-up? 3. What is the shape of a DNA molecule like? Describe it. 4. How does DNA condense from chromatin form during interphase to the shortened and thickened chromosomes found in metaphase? LG #2: The Cell Cy ...
... 2. Identify the structures of adenine, guanine, thymine, and cytosine. Which nucleotides pair-up? 3. What is the shape of a DNA molecule like? Describe it. 4. How does DNA condense from chromatin form during interphase to the shortened and thickened chromosomes found in metaphase? LG #2: The Cell Cy ...
document
... • You can change the way an organism looks or behaves! • This process of taking DNA from one organism and putting into another is called transformation. ...
... • You can change the way an organism looks or behaves! • This process of taking DNA from one organism and putting into another is called transformation. ...
DNA Technology – Mapping a plasmid A first step in working with
... A first step in working with DNA is mapping the DNA molecule. One way to do this is to use restriction enzymes (restriction endonucleases) that are naturally found in bacteria to cut the DNA molecule into fragments, and then perform a gel electrophoresis on the treated DNA. The fragments of DNA can ...
... A first step in working with DNA is mapping the DNA molecule. One way to do this is to use restriction enzymes (restriction endonucleases) that are naturally found in bacteria to cut the DNA molecule into fragments, and then perform a gel electrophoresis on the treated DNA. The fragments of DNA can ...
Replication of plasmids with the p15A origin in Shewanella
... or manganese by Alteromonas putrefaciens. Applied and Environmental Microbiology 55, 700–706. McMurry, L., Petrucci, R.E. Jr. and Levy, S.B. (1980) Active efflux of tetracycline encoded by four genetically different tetracycline resistance determinants in Escherichia coli. Proceedings of the Nationa ...
... or manganese by Alteromonas putrefaciens. Applied and Environmental Microbiology 55, 700–706. McMurry, L., Petrucci, R.E. Jr. and Levy, S.B. (1980) Active efflux of tetracycline encoded by four genetically different tetracycline resistance determinants in Escherichia coli. Proceedings of the Nationa ...
Gel Electrophoresis!
... The use of a vector (usually a virus) to insert a working gene into a cell with a defective version of that gene 1. engineer virus to contain healthy gene 2. Infect patient’s bone marrow/stem cells in lab 3. Inject recombinant stem cells into patient’s bone marrow. – Still in its trial stages, but h ...
... The use of a vector (usually a virus) to insert a working gene into a cell with a defective version of that gene 1. engineer virus to contain healthy gene 2. Infect patient’s bone marrow/stem cells in lab 3. Inject recombinant stem cells into patient’s bone marrow. – Still in its trial stages, but h ...
1 PROTOCOLS FOR LIGATION-INDEPENDENT CLONING
... with a high fidelity DNA polymerase, such as Phusion Hot Start II High Fidelity DNA Polymerase, but omit the final extension step. An enhancer such as 5% DMSO can be added to increase robustness. An example thermal cycling program for Phusion: 98 °C 3 min, (98 °C 30 s 63 °C 30 s 72 °C 45 s/kb) x 35 ...
... with a high fidelity DNA polymerase, such as Phusion Hot Start II High Fidelity DNA Polymerase, but omit the final extension step. An enhancer such as 5% DMSO can be added to increase robustness. An example thermal cycling program for Phusion: 98 °C 3 min, (98 °C 30 s 63 °C 30 s 72 °C 45 s/kb) x 35 ...
Cloning Restriction Fragments of Cellular DNA
... • Others, like HaeIII, cut both strands in the same location yielding "blunt ends" on the restriction fragments. ...
... • Others, like HaeIII, cut both strands in the same location yielding "blunt ends" on the restriction fragments. ...
Genetic Engineering
... A group led by Melvin Simon modified an endogenous circular plasmid in E. coli, the fertility (F) factor present at one or two copies per cell, to create a cloning vector. In reference to its yeast cousin, they called it bacterial artificial chromosome (BAC). With a cloning capacity of 300 kb, BACs ...
... A group led by Melvin Simon modified an endogenous circular plasmid in E. coli, the fertility (F) factor present at one or two copies per cell, to create a cloning vector. In reference to its yeast cousin, they called it bacterial artificial chromosome (BAC). With a cloning capacity of 300 kb, BACs ...
Mobile genetic elements and horizontal gene transfer
... coupled cytoplasmic membrane DNA translocation complex to import the single stranded uptake DNA into cytoplasm. The cytoplasmic membrane DNA translocation complex includes DNA receptor protein, channel protein and ATP-binding protein [15]. The imported single stranded DNA can be integrated into the ...
... coupled cytoplasmic membrane DNA translocation complex to import the single stranded uptake DNA into cytoplasm. The cytoplasmic membrane DNA translocation complex includes DNA receptor protein, channel protein and ATP-binding protein [15]. The imported single stranded DNA can be integrated into the ...
- Flintbox
... Staphylococcus Aureus Strain with GFP/YFP/mCherry Fluorescence Technology Primer: An estimated 32% of the US population is colonized with Staphylococcus aureus, with the highest prevalence among children. The proportion of strains that is methicillin resistant is increasing, with 64% of screened iso ...
... Staphylococcus Aureus Strain with GFP/YFP/mCherry Fluorescence Technology Primer: An estimated 32% of the US population is colonized with Staphylococcus aureus, with the highest prevalence among children. The proportion of strains that is methicillin resistant is increasing, with 64% of screened iso ...
Plasmid
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A plasmid is a small DNA molecule within a cell that is physically separated from a chromosomal DNA and can replicate independently. They are most commonly found in bacteria as small, circular, double-stranded DNA molecules; however, plasmids are sometimes present in archaea and eukaryotic organisms. In nature, plasmids often carry genes that may benefit the survival of the organism, for example antibiotic resistance. While the chromosomes are big and contain all the essential information for living, plasmids usually are very small and contain only additional information. Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms.Plasmids are considered replicons, a unit of DNA capable of replicating autonomously within a suitable host. However, plasmids, like viruses, are not generally classified as life. Plasmids can be transmitted from one bacterium to another (even of another species) via three main mechanisms: transformation, transduction, and conjugation. This host-to-host transfer of genetic material is called horizontal gene transfer, and plasmids can be considered part of the mobilome. Unlike viruses (which encase their genetic material in a protective protein coat called a capsid), plasmids are ""naked"" DNA and do not encode genes necessary to encase the genetic material for transfer to a new host. However, some classes of plasmids encode the conjugative ""sex"" pilus necessary for their own transfer. The size of the plasmid varies from 1 to over 200 kbp, and the number of identical plasmids in a single cell can range anywhere from one to thousands under some circumstances.The relationship between microbes and plasmid DNA is neither parasitic nor mutualistic, because each implies the presence of an independent species living in a detrimental or commensal state with the host organism. Rather, plasmids provide a mechanism for horizontal gene transfer within a population of microbes and typically provide a selective advantage under a given environmental state. Plasmids may carry genes that provide resistance to naturally occurring antibiotics in a competitive environmental niche, or the proteins produced may act as toxins under similar circumstances, or allow the organism to utilize particular organic compounds that would be advantageous when nutrients are scarce.