New and Redesigned pRS Plasmid Shuttle Vectors for Genetic
... screening, and a range of yeast-selectable markers. These markers include the S. cerevisiae biosynthetic genes TRP1, LEU2, and URA3, which can be used with almost all commonly encountered laboratory strains that are auxotrophic for tryptophan, leucine, or uracil, respectively. However, there are lim ...
... screening, and a range of yeast-selectable markers. These markers include the S. cerevisiae biosynthetic genes TRP1, LEU2, and URA3, which can be used with almost all commonly encountered laboratory strains that are auxotrophic for tryptophan, leucine, or uracil, respectively. However, there are lim ...
Cloning of PCR products into TOPO TA vectors
... Introduction: Bacteria (such as E. coli) and yeasts, often contain extrachromosomal DNA molecules called plasmids. Plasmids are physically independent from chromosomes and replicate using their own replication origins and replicative gene products (proteins and RNAs). They often carry genes that enc ...
... Introduction: Bacteria (such as E. coli) and yeasts, often contain extrachromosomal DNA molecules called plasmids. Plasmids are physically independent from chromosomes and replicate using their own replication origins and replicative gene products (proteins and RNAs). They often carry genes that enc ...
Phenotypic Stability of trp Operon Recombinant
... pSC101-trp and RSF2124-trp were originally in E. coli C600-1. Plasmid preparatiorr. Escherichia coli carrying RSF2124-trp was grown ovei night in PBB and diluted 10fold in 100 ml M-9 medium (Clowes & Hayes, 1968) supplemented with Casamino acids (20 g 1-lj and thiamin (1Opg ml-l) or thymine (1Opg ml ...
... pSC101-trp and RSF2124-trp were originally in E. coli C600-1. Plasmid preparatiorr. Escherichia coli carrying RSF2124-trp was grown ovei night in PBB and diluted 10fold in 100 ml M-9 medium (Clowes & Hayes, 1968) supplemented with Casamino acids (20 g 1-lj and thiamin (1Opg ml-l) or thymine (1Opg ml ...
Manipulating DNA - tools and techniques 2012
... The gene is copied and placed into a bacterial plasmid The plasmid is inserted into the bacterial host cell Inside bacterial host cell the plasmid and the gene make twenty copies of itself The bacterial cell copies itself every twenty minutes by ...
... The gene is copied and placed into a bacterial plasmid The plasmid is inserted into the bacterial host cell Inside bacterial host cell the plasmid and the gene make twenty copies of itself The bacterial cell copies itself every twenty minutes by ...
Solutions to 7.014 Problem Set 7
... e) Why are real populations in nature not usually found at Hardy-Weinberg equilibrium? The Hardy-Weinberg equation assumes that all of the following are true: 1. Infinite population size 2. Random mating occurs 3. No selective pressures are present 4. No mutations 5. No migration into or out of a po ...
... e) Why are real populations in nature not usually found at Hardy-Weinberg equilibrium? The Hardy-Weinberg equation assumes that all of the following are true: 1. Infinite population size 2. Random mating occurs 3. No selective pressures are present 4. No mutations 5. No migration into or out of a po ...
Enteric bacteria as model systems
... These days, complete genome sequences are available, so we need only determine a small region of sequence adjacent to our insertion, and gather the remaining information from the published genome sequence. This can be accomplished by cloning, or by PCR-based methods. Alternatively, we can someti ...
... These days, complete genome sequences are available, so we need only determine a small region of sequence adjacent to our insertion, and gather the remaining information from the published genome sequence. This can be accomplished by cloning, or by PCR-based methods. Alternatively, we can someti ...
In the „restriction endonucleases”
... advantage that it is easy and cheap to sustain, it is growing very fast (1 division in every 20 minutes) and its genetic map is well-known. Besides, Escherichia coli, many other prokaryotic and eukaryotic host cells are in use. ...
... advantage that it is easy and cheap to sustain, it is growing very fast (1 division in every 20 minutes) and its genetic map is well-known. Besides, Escherichia coli, many other prokaryotic and eukaryotic host cells are in use. ...
The QIAexpressionist™
... propagating pQE plasmids. These strains can also be used as expression hosts for expressing nontoxic proteins, but they may be less efficient than the M15[pREP4] strain, and expression is regulated less tightly than in strains harboring the pREP4 plasmid. If the expressed protein is toxic to the cel ...
... propagating pQE plasmids. These strains can also be used as expression hosts for expressing nontoxic proteins, but they may be less efficient than the M15[pREP4] strain, and expression is regulated less tightly than in strains harboring the pREP4 plasmid. If the expressed protein is toxic to the cel ...
Teacher`s Guide- labs, worksheets, prelab notes, tests, rubrics
... c. protein- polymer made of amino acids d. gene expression- the translation of a gene into a protein product e. genotype- the genetic makeup of an organism f. phenotype- the outward expression of genes of an organism 2. To genetically transform an entire organism, you must insert the new gene into e ...
... c. protein- polymer made of amino acids d. gene expression- the translation of a gene into a protein product e. genotype- the genetic makeup of an organism f. phenotype- the outward expression of genes of an organism 2. To genetically transform an entire organism, you must insert the new gene into e ...
gal
... …non-virulent units that are inserted in the host chromosome, and multiply via binary fission along with the host DNA, …prophage can re-enter the lytic cycle to complete the virus life cycle. ...
... …non-virulent units that are inserted in the host chromosome, and multiply via binary fission along with the host DNA, …prophage can re-enter the lytic cycle to complete the virus life cycle. ...
Genetic Engineering
... In order to obtain large quantities of the transformed bacteria which have grown and formed colonies, scientists take a sample and put it to grow in another culture medium containing nutrients. The genetic material the bacteria have produced then needs to be isolated, i.e. it has to be separated fro ...
... In order to obtain large quantities of the transformed bacteria which have grown and formed colonies, scientists take a sample and put it to grow in another culture medium containing nutrients. The genetic material the bacteria have produced then needs to be isolated, i.e. it has to be separated fro ...
Bio 102 Practice Problems
... 8. A Bio 102 student gets so excited about the tyrosinase experiment that she decides to try to clone the tyrosinase gene. She grinds up some potato, extracts the DNA from it and digests the DNA with two different restriction enzymes (separately, not together): EcoRI and BamHI. She then obtains a c ...
... 8. A Bio 102 student gets so excited about the tyrosinase experiment that she decides to try to clone the tyrosinase gene. She grinds up some potato, extracts the DNA from it and digests the DNA with two different restriction enzymes (separately, not together): EcoRI and BamHI. She then obtains a c ...
Recombinant DNA Technology (Lecture 13)
... •Small, autonomously replicating DNA molecules •Plasmids are circular DNA molecules of 1-200 kb found in bacteria or yeast cells. They can be considered molecular parasites but they often benefit their host (e.g. provide antibiotic resistance) •Plasmids used for cloning can replicate up to ~3,000 co ...
... •Small, autonomously replicating DNA molecules •Plasmids are circular DNA molecules of 1-200 kb found in bacteria or yeast cells. They can be considered molecular parasites but they often benefit their host (e.g. provide antibiotic resistance) •Plasmids used for cloning can replicate up to ~3,000 co ...
BIOTECHNOLOGY - Bishop Amat Memorial High School
... Select and isolate specific sequence of DNA in organism of choice (ie: human gene) Incorporate selected DNA sequence into DNA of a vector organism Grow and amplify resulting recombinant DNA in a host that reproduces quickly ...
... Select and isolate specific sequence of DNA in organism of choice (ie: human gene) Incorporate selected DNA sequence into DNA of a vector organism Grow and amplify resulting recombinant DNA in a host that reproduces quickly ...
ap ch 18 virus bacteria - Pregitzersninjascienceclasses
... “Resistance” plasmids - contain genes that code for resistance to antibiotics When a bacterial population is exposed to antibiotics, any “sensitive” bacteria are killed by the antibiotics Bacteria that contain the R plasmid with resistance to the antibiotic survive These bacteria then reprod ...
... “Resistance” plasmids - contain genes that code for resistance to antibiotics When a bacterial population is exposed to antibiotics, any “sensitive” bacteria are killed by the antibiotics Bacteria that contain the R plasmid with resistance to the antibiotic survive These bacteria then reprod ...
AP genetic technology
... • DNA is placed at one end of a gel • A current is applied to the gel • DNA molecules are negatively charged and move toward positive end of gel • Smaller molecules move faster than larger ...
... • DNA is placed at one end of a gel • A current is applied to the gel • DNA molecules are negatively charged and move toward positive end of gel • Smaller molecules move faster than larger ...
Genetic Engineering
... from another are fused • The resulting cell divides like a normal embryo ...
... from another are fused • The resulting cell divides like a normal embryo ...
doc Feb 8th, 2010 notes
... Bacteriophage is a virus capable of infecting bacteria. For example, a bacteriophage (48, 502 bp) can infect E.Coli. o Bacteriophages, like plasmid, can be used as vectors and are capable of prolific replication within a cell. One third of its genome is not required for lytic growth, and can be repl ...
... Bacteriophage is a virus capable of infecting bacteria. For example, a bacteriophage (48, 502 bp) can infect E.Coli. o Bacteriophages, like plasmid, can be used as vectors and are capable of prolific replication within a cell. One third of its genome is not required for lytic growth, and can be repl ...
Genes without frontiers?
... evolution (Maynard Smith et al, 1991; Campbell, 2000; Ochman et al, 2000; Gogarten et al, 2002). This evolution need not be slow. The intense selection pressure imposed on microbial communities by worldwide antibiotic use reveals that new multiresistance plasmids can arise from diverse origins and s ...
... evolution (Maynard Smith et al, 1991; Campbell, 2000; Ochman et al, 2000; Gogarten et al, 2002). This evolution need not be slow. The intense selection pressure imposed on microbial communities by worldwide antibiotic use reveals that new multiresistance plasmids can arise from diverse origins and s ...
Appendix: Fusion Gene Plasmid Construction
... was isolated from the pGEM-BAC 4.8 plasmid (3) as a BamH I - Bgl II fragment and subcloned into the BamH I - Bgl II digested -911 IGRP-CAT plasmid in the same orientation. The resulting plasmid contains IGRP promoter sequence from -1342 to +3, with a native IGRP Xba I restriction endonuclease site a ...
... was isolated from the pGEM-BAC 4.8 plasmid (3) as a BamH I - Bgl II fragment and subcloned into the BamH I - Bgl II digested -911 IGRP-CAT plasmid in the same orientation. The resulting plasmid contains IGRP promoter sequence from -1342 to +3, with a native IGRP Xba I restriction endonuclease site a ...
Bacteria Transformation
... enzymes are used to cut the DNA of the plasmid at designated places in the base sequence. The same enzymes are used to cut a section of human DNA. This section of human DNA is then placed into the space in the cut DNA of the bacterial plasmid. The human DNA codes for the synthesis of a product such ...
... enzymes are used to cut the DNA of the plasmid at designated places in the base sequence. The same enzymes are used to cut a section of human DNA. This section of human DNA is then placed into the space in the cut DNA of the bacterial plasmid. The human DNA codes for the synthesis of a product such ...
Stamm revision
... O’Keefe and Beggs, Page 9 whether there is synthetic enhancement/lethality between the two genes (Figure 2). If one of the mutations causes a growth phenotype by itself, such as cold- or heat-sensitivity, suppression of the defect by the second mutation is also possible. 3.2.1 Construction of doubl ...
... O’Keefe and Beggs, Page 9 whether there is synthetic enhancement/lethality between the two genes (Figure 2). If one of the mutations causes a growth phenotype by itself, such as cold- or heat-sensitivity, suppression of the defect by the second mutation is also possible. 3.2.1 Construction of doubl ...
Product Datasheets
... through a PCR cleanup kit. After preparing your DNA fragments by PCR, verify the PCR products by gel electrophoresis. If multiple bands are obtained, gel purify your DNA insert fragment. Be sure to elute the DNA fragments from column using water. ✔ When gel purifying DNA fragments, employ extra caut ...
... through a PCR cleanup kit. After preparing your DNA fragments by PCR, verify the PCR products by gel electrophoresis. If multiple bands are obtained, gel purify your DNA insert fragment. Be sure to elute the DNA fragments from column using water. ✔ When gel purifying DNA fragments, employ extra caut ...
Designing and making sgRNA constructs
... (NOTE: many protocols call for phosphatasing the oligonucleotides. This won't hurt, but it doesn't matter since the vector ends are phosphorylated. Also, some people gel purify the plasmid after digestion. It can't hurt, but if your digest went to completion, this is a waste of time. Essentially, th ...
... (NOTE: many protocols call for phosphatasing the oligonucleotides. This won't hurt, but it doesn't matter since the vector ends are phosphorylated. Also, some people gel purify the plasmid after digestion. It can't hurt, but if your digest went to completion, this is a waste of time. Essentially, th ...
Cloning and functional analysis of
... debris by centrifugation at 15,000 rpm for 30 min at 4°C, the supernatant was applied directly to His-bind resin (Novagen Inc., Darmstadt, Germany). The resin was washed with 20 mM of imidazole and eluted using 200 mM of imidazole in buffer A. To remove imidazole, the purified fraction was passed t ...
... debris by centrifugation at 15,000 rpm for 30 min at 4°C, the supernatant was applied directly to His-bind resin (Novagen Inc., Darmstadt, Germany). The resin was washed with 20 mM of imidazole and eluted using 200 mM of imidazole in buffer A. To remove imidazole, the purified fraction was passed t ...
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.