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Isolation of clonable DNA (genomic libraries, cDNA synthesis) cloning vectors (plasmids, bacteriophages, cosmids) plasmid incompatibility and control of copy number. (up.ac.za)
Use to confirm the function of the products of genes or cDNAs that are thought to encode a phytoene synthase enzyme, or to screen genomic or cDNA libraries for plasmids containing such genes (crtB) or cDNAs (psy). (addgene.org)
In contrast, complementary DNA (cDNA) libraries contain only those DNA sequences that are complementary to messenger RNA (mRNA) molecules present in a cell and differ from one cell type to another. (pharmacy180.com)
Immunization with rTcSSP4 protein makes mice more susceptible to trypomastigote infection, with high mortality rates, whereas mice immunized with a eukaryotic expression plasmid containing the TcSSP4 cDNA were able to control the acute phase of infection. (ijbs.com)

and reintroduced into the bacterium, which will replicate, thus making multiple copies of the hybrid plasmid. (pharmacy180.com)
You have successfully inserted your favorite gene into the cloning vector, and you are ready to make copies of the hybrid plasmid. (quartzy.com)

By removing the genes in the plasmid that caused the tumor and adding in novel genes, researchers were able to infect plants with A. tumefaciens and let the bacteria insert their chosen DNA into the genomes of the plants. (wikipedia.org)
This powerful strategy generates libraries of repetitive genes over a wide range of molecular weights in a 'one-pot' parallel format. (bionity.com)
These libraries provide researchers with a valuable resource for studying genes, their regulatory elements, and their functions. (axis-shield-density-gradient-media.com)
One of the hallmarks of the emergence of M. ulcerans as a more severe pathogen is the acquisition of a 174-kb plasmid that bears a cluster of genes necessary for the synthesis of the polyketide toxin mycolactone. (cdc.gov)
Immunization of mice with plasmids containing genes of Trypanosoma cruzi induces protective immunity in the murine model of Chagas disease. (ijbs.com)
CRISPR has been used for gene editing by inserting a plasmid containing genes and defined repeats, allowing an organism's genome to be cut at any desired location. (columbia.edu)
Eventually, newly-born mosquitos would carry both copies of the resistant genes and this would then spread in the wild. (columbia.edu)

A very popular method for making tandem copies of DNA sequences involves inserting them iteratively into a bacterial plasmid," Amiram said. (bionity.com)
In this example, the human insulin gene is inserted into a bacterial plasmid. (pressbooks.pub)

Plasmids, discovered in 1952, became important tools for transferring information between cells and replicating DNA sequences. (wikipedia.org)
Vertical red lines indicate scaffold breaks for Pav sequences or boundaries between chromosomes/plasmids in the case of the Pto DC3000 reference sequence. (sirnalibrary.com)
Make K copies of each completely "de-ambigulated" variant: E.g. the sequence "SATC" will then be treated as 2 sequences: "GATC" and "CTAC" that will each appear in K copies. (pypi.org)

PCR and PacBio-based whole genome sequencing were used to determine if the plasmid was maintained within the chromosome or as an episome. (oregonstate.edu)
The total genome sizes were 6.6 megabases (Mb) for Pav BP631 and 6.1 to 6.2 Mb for the other two strains, consistent with the presence of extra-chromosomal plasmids in Pav BP631. (sirnalibrary.com)
It ideally contains a copy of every DNA nucleotide sequence in the genome. (pharmacy180.com)
In molecular cloning, a target DNA fragment is inserted into a vector, often a plasmid or a viral genome, using specialized enzymes like restriction endonucleases and DNA ligase. (axis-shield-density-gradient-media.com)
Furthermore, molecular cloning serves as the foundation for constructing genomic libraries, which are comprehensive collections of cloned DNA fragments representing an organism's entire genome. (axis-shield-density-gradient-media.com)
However, by using a prototype plasmid-based microarray that covered 10% of the genome, we found multiple genomic DNA deletions among 30 M. ulcerans clinical isolates of diverse geographic origins. (cdc.gov)
To facilitate M. grisea genome analysis, physical mapping, and positional cloning, we have constructed a bacterial artificial chromosome (BAC) library from the rice infecting strain 70-15. (johnshopkins.edu)
These data show that the BAC library is suitable for genome analysis of M. grisea. (johnshopkins.edu)
Title : A Human Papillomavirus Whole Genome Plasmid Repository: A Resource for HPV DNA Quality Control Reagents Personal Author(s) : Thapa, Hem R.;Unger, Elizabeth R.;Querec, Troy D. (cdc.gov)

To test various conditions that could improve peanut transformation, peanut cultivar Georgia Green was co-bombarded with two plasmids, one containing a green fluorescent protein (GFP) gene and a second containing a gene of interest plus selectable marker. (usda.gov)
The insulin gene from humans was inserted into a plasmid. (pressbooks.pub)
Gain even greater insights into complex biological networks using Sigma-Aldrich ® Advanced Genomics CRISPR activation and CRISPR inhibition libraries for gene knockdown and overexpression experiments. (sigmaaldrich.com)
Therefore, if a single gene from a certain gene family is mutated, there is a high probability that another gene from the same family (actually a copy very similar to the mutated gene) will mask the phenotypes in place of the mutated gene. (phys.org)
The library was used to identify BAC clones containing a adenylate cyclase gene (mac1). (johnshopkins.edu)
Or perhaps you've done some DNA cloning , trying to 'paste' a gene into a circular DNA plasmid. (khanacademy.org)
Now, you want to check and see whether your PCR worked, or whether your plasmid has the right gene in it. (khanacademy.org)
The mcr-1 gene was first reported in 2015 in food, animal, and patient isolates from China ( 1 ) and is notable for being the first plasmid-mediated colistin resistance mechanism to be identified. (cdc.gov)
Plasmids can be transferred between bacteria, potentially spreading the resistance gene to other bacterial species. (cdc.gov)

In the early 1970s it was found that this bacteria inserted its DNA into plants using a Ti plasmid. (wikipedia.org)
This recombinant DNA plasmid was then inserted into bacteria. (pressbooks.pub)
This recombinant plasmid can then be used to transform bacteria, which gain the ability to produce the insulin protein. (pressbooks.pub)
Overnight-grown bacteria were selleck screening library subcultured and grown to exponential phase, harvested by centrifugation (20 min/2700 × g) and resuspended in PBS. (screeninglibraries.com)
In Escherichia coli and related bacteria, we believe that both plasmids and the bacterial chromosome use site-specific recombination to convert multimers to monomers. (gla.ac.uk)

Garvin L, Vande Voorde R, Dickinson M, Carrell S, Hybiske K, Rockey D (2021) A broad-spectrum cloning vector that exists as both an integrated element and a free plasmid in Chlamydia trachomatis. (oregonstate.edu)
After the vector has grown in size, the copies of the sequence are cut out using enzymes and the process is repeated to generate a larger polymer. (bionity.com)
adapt to low pH of M. bovis BCG, containing the empty cloning vector pMV261 [BCG (pMV261)], EVP4593 chemical structure and of M. bovis BCG with the MDP1-antisense-plasmid pAS-MDP1 [BCG (pAS-MDP1)], by comparing the growth without and with pH stress. (sirnalibrary.com)
A genomic library is a collection of fragments of dsDNA obtained by digestion of the total DNA of the organism with a restriction endonuclease and subsequent ligation to an appropriate vector. (pharmacy180.com)

While some plasmids were fully integrated, some were maintained as episomes and could be purified and retransformed into E. coli. (oregonstate.edu)
For plasmid selection and maintenance in E. coli, use chloramphenicol at 30 mg/liter. (addgene.org)
This is achieved by introducing the plasmid into E. coli cells-a process known as "transformation," indicating that the bacterial cells have been genetically modified (i.e. "transformed" by the uptake of the foreign DNA). (quartzy.com)

Sigma-Aldrich ® Advanced Genomics offers multiple Cas9 variants (wild-type, enhanced specificity, nickase, GFP-fused, catalytically inactive) and in several formats (plasmid, lentivirus, lyophilized protein). (sigmaaldrich.com)
It was found that 45-bp deletion abolished binding of a plasmid replication protein to the plasmid DNA, supporting an iteron model for plasmid replication. (readabstracts.com)
The technique involves replicating the DNA coding for a protein into a plasmid. (alternativemedicine.com)
This web-application allows small to medium-sized protein production groups to track data related to plasmid DNA, conditioned media samples (supes), cell lines used for expression, and purified protein information, including method of purification and quality control results. (biomedcentral.com)

We explored the utility of a broad host-range plasmid, pBBR1MCS-4, to transform chlamydiae, with a goal of simplifying the transformation process. (oregonstate.edu)
You then transform this library into an organism that doesnt natively methylate DNA. (pypi.org)
There are two methods to transform competent cells with plasmid DNA: chemical transformation and electroporation. (quartzy.com)

More specifically, you will find information on the plasmid strains we used, the modifications we did to them in order to be compatible with the Biobrick library and our cloning plan. (igem.org)

Results showed that the strain with 13 copies produced 1.78-fold lycopene titer that of the controlled strain with IUP-harbored plasmid, and it exhibited stable expression after ten successions while the plasmid loss was observed in the controlled strain in the 3rd succession. (authorea.com)
However, multimerisation also interferes with the stable inheritance of high copy number plasmids. (gla.ac.uk)

PCR permits the synthesis of millions of copies of a specific nucleotide sequence in a few hours. (pharmacy180.com)

PacBio sequencing of the cloned transformants revealed allelic exchange events between the chromosome and plasmid pBVR2 that replaced chromosomal incA with the plasmid GFP:CAT cassette. (oregonstate.edu)
The data also showed evidence of full integration of the plasmid into the bacterial chromosome. (oregonstate.edu)

Grow, Harvest, Sequence plasmids. (pypi.org)

Plasmid transformation requires a replicon derived from the native Chlamydia plasmid, and these transformations are species-specific. (oregonstate.edu)
Clearly, a unit copy replicon in dimeric form can not be normally partitioned into two daughter cells. (gla.ac.uk)

Prokaryotic organisms normally contain small, circular, extrachromosomal DNA molecules called plasmids that can serve as vectors. (pharmacy180.com)

As to effectiveness, promotor engineering strategy was employed to regulate IUP expression level, while ribosome-binding site (RBS) library of the key enzyme was constructed for screening the optimal RBS, followed by optimization of concentration of inducer and substrates, the titer of reporting production, lycopene, from 0.087 to 8.67 mg/OD600. (authorea.com)

The two fragments were ligated together to give plasmid pAC-85b. (addgene.org)
A DNA library is a collection of cloned restriction fragments of the DNA of an organism. (pharmacy180.com)
A new method was developed for separation of partially digested large-molecular-weight DNA fragments that facilitated library construction with large inserts. (johnshopkins.edu)

Once the gel is in the box, each of the DNA samples we want to examine (for instance, each PCR reaction or each restriction-digested plasmid) is carefully transferred into one of the wells. (khanacademy.org)

Plasmid pAC-BETA was digested with EcoRI and partially digested with BamHI and a 7.1 kB fragment was gel-purified. (addgene.org)

Unfortunately, working with low copy plasmids makes the procedure more demanding in the lab. (igem.org)

You then clone this cassette into a plasmid with 1 DNA MTase in each plasmid. (pypi.org)

The DNA coating agent protamine was shown to improve transformation efficiency in rice, while a reduction of plasmid DNA in the bombardment mixture was reported to elevate the chance of recovering single copy integration events. (usda.gov)
A 4.6-fold improvement of transformation efficiency was achieved in stably transformed peanut lines by introducing protamine instead of conventional spermidine in a bombardment mixture with 70 ng/shot of plasmid DNA and 50 µg/shot of gold. (usda.gov)
It is the best choice for applications such as construction of libraries that require the best possible transformation efficiencies. (quartzy.com)

The use of Red/ET recombination to generate a large number of shuttle plasmids from a diverse library of chimeric domains for directed evolution of the pikromycin polyketide synthase (PKS) is reported. (readabstracts.com)

A mechanism by which SCP2 derived plasmid achieves increased levels of metabolite production was identified to examine the 45-bp deletion mutation in the plasmid replication origin increased plasmid copy number. (readabstracts.com)

This should help us to make large libraries of proteins, which we can use to rapidly screen new combinations. (bionity.com)
It is important to insert parts responsible for the production of fluorescent proteins in low copy plasmids, as they are potentially harmful for the cell. (igem.org)
ProteinTracker makes extensive use of existing open-source libraries and is designed to track essential data related to the production and purification of proteins. (biomedcentral.com)

Plasmid pACYC184 was digested with EcoRI and BamHI and a 2.4 kB fragment was gel-purified. (addgene.org)
The term "cloning" refers to the replication of a specific DNA sequence or an entire organism, and in the molecular biology context, it primarily involves the generation of identical copies of a DNA fragment. (axis-shield-density-gradient-media.com)

The final plasmid construct, pBVR2, was successfully transformed into C. trachomatis strain L2-434. (oregonstate.edu)
Using a combinatorial pathway library approach, we first mined a historical industrial strain collection for superior enzymes that we used to engineer the autotrophic acetogen Clostridium autoethanogenum . (nature.com)
These high-coverage contigs indicate that this strain harbors one or more multi-copy plasmids. (sirnalibrary.com)

For better yield of low copy number pAC-based plasmids, grow liquid cultures on a platform shaker at ca. 30 degrees Celsius. (addgene.org)

We identified SMT4 , which encodes a SUMO isopeptidase, as a high copy suppressor of both the temperature sensitivity and precocious sister dissociation of pds5 mutants. (silverchair.com)

We have also made a number of improvements to Inventory, added locking to embedded Tables in Worksheets, improved the process for adding multiple Internal References, enhanced our Materials Libraries and extended our External APIs. (perkinelmer.com)

Probes can be used to identify which clone of a library or which band on a gel contains the target DNA. (pharmacy180.com)

Given its speed and high-throughput nature, qPCR can be employed as an effective screening tool to separate high copy number events from low copy events as early as the shoot formation stage of regeneration. (usda.gov)
Many of the changes appear to have been mediated by insertion sequence (IS) elements IS 2404 and IS 2606 , which have high copy numbers. (cdc.gov)

A correlation of 95% was found for estimation of copy number between Southern blot and q-PCR data. (usda.gov)
Data is a raw and unorganized fact that required to be processed to make it In the movie, the scene takes place in the library of the National warzone 2 unlock tool script of Health. (winsomesourcing.com)
Managing reagent data stored in files is challenging because files are often copied, moved, and reformatted. (biomedcentral.com)

In order to determine the transgene copy number during plantlet regeneration, relative quantitative realtime-PCR (qPCR) was established using fluorescently labeled universal library probes. (usda.gov)

Introduction of a foreign DNA molecule into a replicating cell permits the amplification (production of many copies) of the DNA, a process called cloning. (pharmacy180.com)

Administrators are recommended to subscribe to the channels within our support news site found at https://support.revvitysignals.com/hc/en-us/categories/360004446171-Support-News which contains more information about releases and other pertinent product information. (perkinelmer.com)

Unexpectedly, the reduction of plasmid DNA from 700 ng/shot to 70 ng/shot produced transgenic lines with significantly increased number of transgene copies. (usda.gov)

siRNAs and plasmids were used to silence or enhance the expression of NOX4. (jcancer.org)

In EndNote Library, go to File - Import. (jcancer.org)

You may not be able to create an account or request plasmids through this website until you upgrade your browser. (addgene.org)

Moreover, in this page you may find here an (unfortunately not complete) electronic copy of our lab notebook. (igem.org)

Experts can insert the plasmid into an animal or bacterial cells through various processes. (alternativemedicine.com)

For our system we needed three plasmids with different origins of replication and antibiotic resistances. (igem.org)
Corrigendum to "Optimization of a two-plasmid system for the identification of promoters recognized by RNA polymerase containing Staphylococcus aureus alternative sigma factor σB" [FEMS Microbiol. (uzh.ch)

While this pooled approach has enabled rapid generation and screening of complex libraries, successful implementation of pooled screens requires careful choices of experimental parameters. (biomedcentral.com)

Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications. (addgene.org)

The plasmid pAdPSY1 can be used as a positive control. (addgene.org)

Copies of colony hybridization membranes are available upon request. (johnshopkins.edu)

Anatomy7

Organisms24

Diseases2

Chemicals and Drugs60

Analytical, Diagnostic and Therapeutic Techniques and Equipment40

Phenomena and Processes102

Disciplines and Occupations2

Anthropology, Education, Sociology and Social Phenomena3

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Information Science31

Named Groups1

Health Care6

CDNA4

Hybrid plasmid2

Genes7

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Sequences3

Genome9

Gene9

Bacteria5

Vector4

Coli3

Protein4

Transform3

Strains1

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Harvest1

Replicon2

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Partially digested1

Procedure1

Cassette1

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Shuttle1

Mechanism1

Proteins3

Fragment2

Strain3

Grow1

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References1

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High copy2

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