Transform cells and spread plates
... o Observe the color and glow of the bacteria under a UV light o Fewer colonies of bacteria if amp negatively affected growth o Equal numbers of colonies on both LB nutrient agar and LB/amp agar if amp had no effect o The presence of any colonies on the amp plate would suggest that those bacteria are ...
... o Observe the color and glow of the bacteria under a UV light o Fewer colonies of bacteria if amp negatively affected growth o Equal numbers of colonies on both LB nutrient agar and LB/amp agar if amp had no effect o The presence of any colonies on the amp plate would suggest that those bacteria are ...
Supplementary methods
... qRT-PCR Reverse transcription was performed using and random hexamer primer (Fermentas) or a gene specific primer (as indicated) with the M-MuLV reverse transcriptase (Fermentas) as recommended by the manufacturer. qPCR was performed using gene specific pirmers (Table S5) as described in the qChIP m ...
... qRT-PCR Reverse transcription was performed using and random hexamer primer (Fermentas) or a gene specific primer (as indicated) with the M-MuLV reverse transcriptase (Fermentas) as recommended by the manufacturer. qPCR was performed using gene specific pirmers (Table S5) as described in the qChIP m ...
The_RAY_Manual
... E. coli as well as ES-cells, permitting a selection for the recombination product in E.coli. Cotransformed yeast colonies are pooled, extrachromosomal DNA is prepared and electroporated into E. coli. Bacterial transformants containing the recombination product are selected on plates containing kana ...
... E. coli as well as ES-cells, permitting a selection for the recombination product in E.coli. Cotransformed yeast colonies are pooled, extrachromosomal DNA is prepared and electroporated into E. coli. Bacterial transformants containing the recombination product are selected on plates containing kana ...
PCR-based cloning from plasmids Entered by Karin Holmberg
... 4. Excise bands corresponding to the linearized vector and purify with the PureLink Quick Gel Extraction Kit (Invitrogen #K2100-25) according to the manufacturer’s recommendation. ...
... 4. Excise bands corresponding to the linearized vector and purify with the PureLink Quick Gel Extraction Kit (Invitrogen #K2100-25) according to the manufacturer’s recommendation. ...
Chapter 18
... The normal allele of a gene is inserted into a plasmid, with a reporter gene in the middle of the normal allele. The recombinant plasmid transfects mouse embryonic stem cells. The sequences line up with homologous sequences, and if recombination occurs, the normal allele is lost because the plasmid ...
... The normal allele of a gene is inserted into a plasmid, with a reporter gene in the middle of the normal allele. The recombinant plasmid transfects mouse embryonic stem cells. The sequences line up with homologous sequences, and if recombination occurs, the normal allele is lost because the plasmid ...
Biol 101 Study Guide Exam 5
... B) Viruses can enter a host cell when the protein molecules on the outside of the virus fit into receptor molecules on the outside of the cell. C) A virus is generally considered to be alive because it is cellular and can reproduce on its own. D) The host cell provides most of the tools and raw mate ...
... B) Viruses can enter a host cell when the protein molecules on the outside of the virus fit into receptor molecules on the outside of the cell. C) A virus is generally considered to be alive because it is cellular and can reproduce on its own. D) The host cell provides most of the tools and raw mate ...
Allele replacement: an application that permits rapid manipulation of
... for gene replacement in E. coli.18 The procedure requires two reagents, an HSV-BAC and a gene replacement vector. The construction of the first reagent follows standard virological techniques and exploits the fact that the HSV genome circularizes in the nucleus of infected cells.20,21 Other herpesvi ...
... for gene replacement in E. coli.18 The procedure requires two reagents, an HSV-BAC and a gene replacement vector. The construction of the first reagent follows standard virological techniques and exploits the fact that the HSV genome circularizes in the nucleus of infected cells.20,21 Other herpesvi ...
ACEMBL System:
... promoters T7 and Lac, as well as the T7 terminator element (Illustr.1, 10). The T7 system is currently most commonly used; it requires bacterial strains which contain a T7 polymerase gene in the E. coli genome. The Lac promoter is a strong endogenous promoter which can be utilized in most strains. A ...
... promoters T7 and Lac, as well as the T7 terminator element (Illustr.1, 10). The T7 system is currently most commonly used; it requires bacterial strains which contain a T7 polymerase gene in the E. coli genome. The Lac promoter is a strong endogenous promoter which can be utilized in most strains. A ...
Slide 1
... JC3272I, JW1271, JW1272. The genes yciS and yciM can be found within a 1.8 Kbp fragment of the E. coli chromosome. The restriction enzymes BamHI and SmaI will be used to cut the DNA before and after the genes yciS and yciM. The genes will be then inserted into plasmid puc19 and cloned to obtain an a ...
... JC3272I, JW1271, JW1272. The genes yciS and yciM can be found within a 1.8 Kbp fragment of the E. coli chromosome. The restriction enzymes BamHI and SmaI will be used to cut the DNA before and after the genes yciS and yciM. The genes will be then inserted into plasmid puc19 and cloned to obtain an a ...
Modeling Chromosome Maintenance as a Property of Cell Cycle in
... With the advent of genome-level techniques for rapid identification of gene function, it is becoming important to develop rapid methods for generating hypotheses for their mechanisms of action. One way to investigate the mechanisms by which these genes may participate jointly in a common biological ...
... With the advent of genome-level techniques for rapid identification of gene function, it is becoming important to develop rapid methods for generating hypotheses for their mechanisms of action. One way to investigate the mechanisms by which these genes may participate jointly in a common biological ...
The Genetic Code
... (b, 5 pts) Now you ligate the DNA you produced in part (a) to the sequence below, which you have also cut with the same restriction enzyme. Draw the shortest DNA product that could form from ligating a piece of DNA from part (a) to a piece of DNA from part (b). Make sure to draw the nucleotide seque ...
... (b, 5 pts) Now you ligate the DNA you produced in part (a) to the sequence below, which you have also cut with the same restriction enzyme. Draw the shortest DNA product that could form from ligating a piece of DNA from part (a) to a piece of DNA from part (b). Make sure to draw the nucleotide seque ...
Chapter 8: Recombinant DNA Technology 1. Tools of Recombinant
... Harvest copies of gene to insert into plants or animals ...
... Harvest copies of gene to insert into plants or animals ...
Journal of Bacteriology
... Resistance to antimonite [Sb(III)], arsenite [As(III)], and arsenate [As(V)] is encoded by both plasmid-borne and chromosomal arsenical resistance (ars) operons (2, 3, 10). These operons encode transport systems that extrude the toxic metalloids, thus lowering the intracellular concentration and pro ...
... Resistance to antimonite [Sb(III)], arsenite [As(III)], and arsenate [As(V)] is encoded by both plasmid-borne and chromosomal arsenical resistance (ars) operons (2, 3, 10). These operons encode transport systems that extrude the toxic metalloids, thus lowering the intracellular concentration and pro ...
Recombinant DNA Technology
... be separated from E. coli that do not (e.g., antibiotic resistance, grow cells on antibiotic; only those cells with the anti-biotic resistance grow in colony). ...
... be separated from E. coli that do not (e.g., antibiotic resistance, grow cells on antibiotic; only those cells with the anti-biotic resistance grow in colony). ...
Insertion (sufB) in the anticodon loop or base substitution (sufC) in
... more products from the same part of the mRNA, and in regulation of gene expression. The role of tRNA in such non-triplet reading was early established by the isolation of mutant tRNAs able to suppress certain frameshift mutations. The first suppressors of this kind to be characterized were the sufA, ...
... more products from the same part of the mRNA, and in regulation of gene expression. The role of tRNA in such non-triplet reading was early established by the isolation of mutant tRNAs able to suppress certain frameshift mutations. The first suppressors of this kind to be characterized were the sufA, ...
Protocol for Control Reaction (E0554) | NEB
... high-efficiency transformation, enables high numbers of transformants for simple mutagenesis experiments (substitutions and deletions), and more complex strategies (insertions). In contrast, linear amplification methods typically generate fewer colonies in substitution experiments and are unable to ...
... high-efficiency transformation, enables high numbers of transformants for simple mutagenesis experiments (substitutions and deletions), and more complex strategies (insertions). In contrast, linear amplification methods typically generate fewer colonies in substitution experiments and are unable to ...
Objective 2.1 Lesson D Recombinant Organisms
... Cut open your PLASMID at ONE site only (this may or may not be possible depending upon how you constructed your plasmid). The same enzyme should be able to cut your cell DNA at TWO SITES, one above and one below the gene for insulin. It is very important that you find an enzyme cuts as close to ...
... Cut open your PLASMID at ONE site only (this may or may not be possible depending upon how you constructed your plasmid). The same enzyme should be able to cut your cell DNA at TWO SITES, one above and one below the gene for insulin. It is very important that you find an enzyme cuts as close to ...
Welcome
... 4. Restriction digestion & insertion: The same restriction enzyme is used to cleave the plasmid vector as well as the DNA sequence containing the gene of interest at their specific restriction sites. The gene of interest is then inserted into the plasmid which can re-anneal with the gene sequence du ...
... 4. Restriction digestion & insertion: The same restriction enzyme is used to cleave the plasmid vector as well as the DNA sequence containing the gene of interest at their specific restriction sites. The gene of interest is then inserted into the plasmid which can re-anneal with the gene sequence du ...
AS 09 Genetic Engineering.pps237.5 KB
... converted to single stranded DNA by treatment with ....................................... . This is then treated with ................................................... to produce double stranded (double helix) DNA. Plasmid DNA is also extracted from suitable bacteria for use as a ................ ...
... converted to single stranded DNA by treatment with ....................................... . This is then treated with ................................................... to produce double stranded (double helix) DNA. Plasmid DNA is also extracted from suitable bacteria for use as a ................ ...
Protocols - BioMed Central
... d. Heat shock the cells by incubating them at 42 oC for 45 seconds, immediately afterward return the cells to ice and incubate for 2 minutes. e. Add 450 l SOC medium, mix by inverting the tubes a couple of times and incubate for 1 hour at 37 oC with 300 rpm. f. Pellet the cells in a table top centr ...
... d. Heat shock the cells by incubating them at 42 oC for 45 seconds, immediately afterward return the cells to ice and incubate for 2 minutes. e. Add 450 l SOC medium, mix by inverting the tubes a couple of times and incubate for 1 hour at 37 oC with 300 rpm. f. Pellet the cells in a table top centr ...
E. coli - Sonoma Valley High School
... Samples from the restriction enzyme digests are introduced into the gel. Electric current is applied causing fragments to migrate through the gel. ...
... Samples from the restriction enzyme digests are introduced into the gel. Electric current is applied causing fragments to migrate through the gel. ...
l Saccharomyces cerevisiae as a Genetic Model Organism
... or others depending on the genotype of the strain and its ability to utilize various carbon sources. Glucose is the richest and most readily available carbon source and a rich medium containing glucose is referred to as YEPD or YPD. Because of the abundant nutrient supply, cells divide rapidly on a ...
... or others depending on the genotype of the strain and its ability to utilize various carbon sources. Glucose is the richest and most readily available carbon source and a rich medium containing glucose is referred to as YEPD or YPD. Because of the abundant nutrient supply, cells divide rapidly on a ...
Biotechnology
... Selection of Transformants • Hosts are chosen that are sensitive to a particular substance or require a particular nutrient (auxotrophs) • The vector provides the genes needed to be resistant to the substance or produce the nutrient • Host cells taking up vector or recombinant vector live • Host ce ...
... Selection of Transformants • Hosts are chosen that are sensitive to a particular substance or require a particular nutrient (auxotrophs) • The vector provides the genes needed to be resistant to the substance or produce the nutrient • Host cells taking up vector or recombinant vector live • Host ce ...
emboj7601343-sup
... in a 1:1 mix of Hams Nutrient Mixture F12 (Cambrex) and DMEM supplemented with 5% FBS, 10 g/ml insulin (Sigma), 20ng/ml Epidermal Growth Factor (Sigma), 5g/ml hydrocortisone (Sigma) and penicillin/streptomycin/L-glutamine as above. U-2 OS cells containing stably transfected p53 and p52 RNAi or con ...
... in a 1:1 mix of Hams Nutrient Mixture F12 (Cambrex) and DMEM supplemented with 5% FBS, 10 g/ml insulin (Sigma), 20ng/ml Epidermal Growth Factor (Sigma), 5g/ml hydrocortisone (Sigma) and penicillin/streptomycin/L-glutamine as above. U-2 OS cells containing stably transfected p53 and p52 RNAi or con ...
techniques in molecular biology – methods
... TECHNIQUES IN MOLECULAR BIOLOGY – METHODS FOR PLASMID DNA ISOLATION DNA isolation: The application of molecular biology techniques to the analysis of complex genomes depends on the ability to prepare pure plasmid DNA. Most plasmid DNA isolation techniques come in two flavors, simple - low quality DN ...
... TECHNIQUES IN MOLECULAR BIOLOGY – METHODS FOR PLASMID DNA ISOLATION DNA isolation: The application of molecular biology techniques to the analysis of complex genomes depends on the ability to prepare pure plasmid DNA. Most plasmid DNA isolation techniques come in two flavors, simple - low quality DN ...
Plasmid
.svg?width=300)
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.