Miniprep sequence plasmids. Medical search. Frequent questions

The plasmid DNA yield will vary depending on the plasmid copy number, type and size, the bacterial strain, the growth conditions, and the kit. (wikipedia.org)
Up to 100 µg per preparation, depending on the plasmid copy number, culture growth conditions, and strain of E. coli utilized. (zymoresearch.eu)

Many methods have been developed to purify plasmid DNA from bacteria. (wikipedia.org)
Kits are available from varying manufacturers to purify plasmid DNA, which are named by size of bacterial culture and corresponding plasmid yield. (wikipedia.org)
Automation of the protocol on an ASSIST PLUS pipetting robot with a VIAFLO 12 channel 1250 μl electronic pipette offers an easy and efficient way to extract and purify plasmid DNA with minimal hands-on time, guaranteeing perfect and reproducible liquid handling while protecting the user from repetitive strain injuries. (integra-biosciences.com)
The kit has been used to purify plasmid DNA for high efficiency transfection. (sigmaaldrich.com)

The efficacy of pSelAct-KO was demonstrated in C. michiganensis and confirmed using whole genome sequencing. (biorxiv.org)
Although short-read sequencing technologies have enabled large-scale bacterial genomics, the resulting draft genome assemblies are often fragmented into hundreds of discrete contigs. (gointerlabs.com)
The majority of sequences grouped in clade A, which was represented by the mrk gene cluster from the genome sequenced K. pneumoniae MGH78578. (biomedcentral.com)
however, discovery and genotyping of molecular markers in oat has been hindered by the size and complexity of the genome, and by a scarcity of sequence data. (biomedcentral.com)
Advances in sequencing technology have enhanced genome-wide SNP discovery, and SNP platforms have been developed for a number of diploid species. (biomedcentral.com)

A plasmid preparation is a method of DNA extraction and purification for plasmid DNA, it is an important step in many molecular biology experiments and is essential for the successful use of plasmids in research and biotechnology. (wikipedia.org)
During the purification procedure, the plasmid DNA is often separated from contaminating proteins and genomic DNA. (wikipedia.org)
These methods invariably involve three steps: growth of the bacterial culture, harvesting and lysis of the bacteria, and purification of the plasmid DNA. (wikipedia.org)
Purification of plasmids is central to molecular cloning. (wikipedia.org)
It is important to consider the downstream applications of the plasmid DNA when choosing a purification method. (wikipedia.org)
For example, if the plasmid is to be used for transfection or electroporation, a purification method that results in high purity and low endotoxin levels is desirable. (wikipedia.org)
Similarly, if the plasmid is to be used for sequencing or PCR, a purification method that results in high yield and minimal contaminants is desirable. (wikipedia.org)
The use of silica membrane-based DNA purification kits is a convenient way to prepare plasmid DNA samples for cloning or subsequent analysis, e.g. sequencing or restriction analysis. (integra-biosciences.com)
MACHEREY-NAGEL's NucleoSpin 96 Plasmid kit is designed for high throughput purification of high-copy plasmid DNA from E. coli in a 96 well plate format. (integra-biosciences.com)
MACHEREY-NAGEL's NucleoSpin 96 Plasmid kit and NucleoVac 96 Vacuum Manifold are a proven method for high throughput plasmid DNA purification. (integra-biosciences.com)
The ASSIST PLUS pipetting robot is used to automate the pipetting steps of the MACHEREY-NAGEL NucleoSpin 96 Plasmid kit protocol for plasmid DNA purification. (integra-biosciences.com)
For spin-column purification of endotoxin-free BAC/PAC plasmid DNA (up to ~200 kb) for sequencing, PCR, restriction endonuclease digestion, etc. (kyongshin.com)
The ZymoPURE™ 96 Plasmid Miniprep Kit features a high-throughput method for the purification of up to 100 µg of ultra-pure transfection-grade plasmid DNA using a vacuum manifold or centrifuge. (zymoresearch.eu)
ZymoPURE™ technology uses a modified alkaline lysis method and our patented binding chemistry, which enables the highest DNA binding capacity and rapid loading of the lysate and wash buffer, resulting in the purification of highly concentrated (up to 1 µg/µl) plasmid DNA directly from a 96-well vacuum plate. (zymoresearch.eu)

The DNA is ready for immediate use in downstream applications such as transfection, restriction digestion, cloning, sequencing, and PCR amplification. (sigmaaldrich.com)
Brie﫿y, two single-stranded oligonucleotides are annealed to form a double-stranded DNA sequence containing the shRNA sequence followed by a Pol III terminator (consisting of a stretch of four or more thymidines), as well as 5'- and 3'-end "sticky" overhangs that are complementary to those generated in the targeting vector following restriction digestion with BglII and SalI (Figures 3E and 4C). (berterolab.com)
Furthermore, the oligonucleotides are designed to destroy the BglII restriction enzyme site in the 﫿nal plasmid in order to allow screening of correct recombinant clones by diagnostic restriction digestion (Figure 4D). (berterolab.com)

Plasmids are almost always purified from liquid bacteria cultures, usually E. coli, which have been transformed and isolated. (wikipedia.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)
Transformed E.coli 상태로 의뢰시 배양 후 plasmid miniprep을 진행해 드립니다. (cosmogenetech.com)
The E. coli and K. pneumoniae mrkABCD gene sequences clustered together in two distinct clades, supporting previous evidence for the occurrence of inter-genera lateral gene transfer. (biomedcentral.com)
We introduce a simple, dual direct cloning plasmid system (pgMAX-II) for gene expression analysis in both prokaryotic (Escherichia coli) and mammalian cells. (bvsalud.org)

Primers 16S 440F and 16S 1491R (Table) were ing of glycopeptide antimicrobial agents to their designed to amplify partial 16S rDNA sequences. (cdc.gov)
The pCR™4-TOPO™ TA vector has sites for 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3. (thermofisher.com)
Using TOPO™ cloning technology, there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. (thermofisher.com)
Standard primers offered by MRC PPU DNA Sequencing and Services are free! (dnaseq.co.uk)
Through phosphorothioate modification of primers, we realized onepot detection with high sensitivity using plasmids of SARS-CoV-2, HPV16 and HPV18. (gointerlabs.com)
Ogle and TAM amplicons from 12 primers were sequenced for SNP validation, revealing complex polymorphism in seven amplicons but general sequence conservation within SNP loci. (biomedcentral.com)

The GenElute Endotoxin-free Plasmid Kit offers a simple, rapid, cost effective method for purifying plasmid DNA with ≤0.1 EU/μg DNA for high efficiency transfection. (sigmaaldrich.com)
After a simple spin-wash step, the bound plasmid DNA is eluted in endotoxin-free water. (sigmaaldrich.com)
Up to 250 μg of endotoxin-free plasmid DNA may be prepared from 5 to 40 mL of culture. (sigmaaldrich.com)

There are two methods to transform competent cells with plasmid DNA: chemical transformation and electroporation. (quartzy.com)
Overnight inoculation of ISA, ISI, ISD, Streptavidin initial plasmid transformed cells and competent cells. (igem.org)

We also identified several genes that do not appear to be protein coding, and generated 17 kb of novel transcript sequence data from re-sequencing 97 EST clones. (biomedcentral.com)
Prokaryotic expression screening of the ß2 subunit using an epitope library of CaV1.2 resulted in two overlapping clones of the C-terminal sequence of CaV1.2. (bvsalud.org)

Endotoxins (also known as lipopolysaccharides or LPS) are often co-purified with plasmid DNA and significantly reduce transfection efficiencies in endotoxin-sensitive cell lines. (sigmaaldrich.com)
Eluted Plasmid DNA is Transfection-Grade and Free of Cross-Contamination. (zymoresearch.eu)

In addition, QuickLyse technology uses fewer buffers, simplifying handling and saving time: 24 plasmid DNA minipreps can be prepared in less than 22 minutes. (qiagen.com)
Perform 96 Plasmid Minipreps Simultaneously using a Vacuum Manifold or Centrifuge. (zymoresearch.eu)

The isolated DNA can be used directly in standard applications, including automated sequencing, PCR, restriction analysis, and cloning. (qiagen.com)

The QuickLyse Miniprep Kit provides a rapid spin-column-based method for isolating sequencing-grade plasmid DNA. (qiagen.com)
Commonly used miniprep methods include alkaline lysis and spin-column based kits. (wikipedia.org)

As an added convenience, the ZymoPURE™ 96 Plasmid Miniprep Kit contains colored buffers that permit error-free visualization and identification of complete bacterial cell lysis and neutralization. (zymoresearch.eu)

We developed permissR, an R package to identify permissive sites for chromosomal integration, which can be used in conjunction with pSelAct-Express, a non-replicating integrative plasmid that enables recombination into a permissive genomic location. (biorxiv.org)
Author Manuscript genomic DNA sequence of interest that lies next to a protospacer adjacent motif (PAM) matching the sequence NGG5-12, 14 (Supplementary Fig. 1). (cdc.gov)

Virtually all plasmid vectors in common use encode one or more antibiotic resistance genes as a selectable marker, for example a gene encoding ampicillin or kanamycin resistance, which allows bacteria that have been successfully transformed to multiply uninhibited. (wikipedia.org)
Bacteria that have not taken up the plasmid vector are assumed to lack the resistance gene, and thus only colonies representing successful transformations are expected to grow. (wikipedia.org)
Currently around a million experiments are performed simultaneously for gene expression analysis 2 and around a billion for next generation DNA sequencing 3 . (nature.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)
All of the strains examined caused type 3 fimbriae mediated agglutination of tannic acid treated human erythrocytes despite sequence variation in the mrkD -encoding adhesin gene. (biomedcentral.com)
A: Specify what gene or destination plasmid you need and indicate that you don't have source plasmids. (customdnaconstructs.com)

The TOPO™ TA Cloning™ Kits for Sequencing provide a highly efficient, 5 minute, one-step cloning strategy ('TOPO™ Cloning') for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for sequencing. (thermofisher.com)
Each kit uses the pCR™4-TOPO™ TA vector with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction. (thermofisher.com)
We have removed much of the multiple cloning site from the pCR™4-TOPO™ TA vector to shorten the distance between sequencing primer sites and the insert site to as little as 33 bp. (thermofisher.com)
This means sequencing reactions give less vector sequence and more insert sequence. (thermofisher.com)
The 﫿rst step in generating OPTiKD hPSCs consists in obtaining oligonucleotides to be used to clone the shRNA sequence in the targeting vector. (berterolab.com)

Here we propose using the Kraken classifier, together with a custom Kraken database comprising known chromosomal and plasmid sequences of Klebsiella pneumoniae species complex (KpSC), to identify plasmid -derived contigs in draft assemblies. (gointerlabs.com)

Positive amplification of the target sequence resulted in a colour change from pink to yellow. (biomedcentral.com)

A typical plasmid DNA yield of a miniprep is 5 to 50 µg depending on the cell strain. (wikipedia.org)

The QuickLyse Miniprep Kit yields sufficient plasmid DNA for any downstream application, including automated sequencing. (qiagen.com)
A: Plasmids are prepared as miniprep DNA, a small amount, sufficient for multiple transformations, is sent in 10ul TE. (customdnaconstructs.com)

Q5: Can the ZymoPURE kits be used to isolate large plasmid constructs (BAC/PAC)? (zymoresearch.eu)

The alkaline lysis-based miniprep protocol typically yields 5-15 μg of plasmid DNA from 1.5ml overnight cultures. (integra-biosciences.com)

Sequence alignment analysis showed 5-14.8% SNP in the partial sequences of the 18 s rRNA of P. falciparum . (biomedcentral.com)

Furthermore, nucleotide sequences related to the cluster vanHAX are present in this DNA, suggesting that the prolonged use of avoparcin in agriculture led to the uptake of glycopeptide resistance genes by animal commensal bacteria, which were subsequently transferred to humans. (cdc.gov)
Plasmids play an important role in bacterial evolution and mediate horizontal transfer of genes including virulence and antimicrobial resistance genes. (gointerlabs.com)
Genes that are directly regulated by SigB contain a promotor binding motif (PBM) with a previously identified consensus sequence. (biomedcentral.com)
The 255 genes were assigned to five categories (I-V) based on their similarity to the original SigB consensus sequences. (biomedcentral.com)
The rapid emergence of antibiotic resistance among bacteria is, to a great extent, due to the dissemination of antibiotic resistance genes by horizontal transfer mediated by plasmids, transposons and integrons (5). (scielo.br)

SNP assays directly interrogate the sequence variation, reducing genotyping errors compared to assays based on size discrimination or hybridization. (biomedcentral.com)

We perform the cycle sequencing reactions on the day the samples are supplied to us, followed by evaluation of productions on our sequencers. (dnaseq.co.uk)
Beginning of gibson reactions involving the previously amplified DNA sequences. (igem.org)

To increase elution of large plasmid DNA, we recommend pre-warming the ZymoPURE Elution Buffer (50 ºC) and increasing the incubation time on column up to 10 minutes prior to centrifugation. (zymoresearch.eu)

6). Detection of specific nucleic acid sequences by hybridization a. mini-prep of plasmid DNA b. (ntu.edu.tw)

Based on cDNA libraries of four cultivated oat genotypes, approximately 127,000 contigs were assembled from approximately one million Roche 454 sequence reads. (biomedcentral.com)

High yields of high-quality plasmid DNA. (qiagen.com)

일반적인 sequencing 서비스로 plasmid나 PCR product를 지정하신 primer를 이용하여 염기서열 분석을 수행합니다. (cosmogenetech.com)
Targeted sequencing하고자 하실때 제공해 주시거나 또는 당사 PCR optimization service를 통하여 제작된 primer를 사용하여 PCR을 진행해 드리는 서비스입니다. (cosmogenetech.com)
PCR product는 sequencing 반응에 영향을 미칠 수 있는 잔류 dNTPs와 primer를 반드시 제거해주어야 합니다. (cosmogenetech.com)
Sequencing primer 합성 서비스와 연계한 primer walking 법으로 정제된 plasmid DNA 전체 서열을 분석해 드리는 서비스입니다. (cosmogenetech.com)
제공된 정보에 따라 sequencing primer를 합성하고, primer walking방식으로 진행됩니다. (cosmogenetech.com)
Primer 1/3 were employed to check insertion of targeted sequence after TA cloning. (biomedcentral.com)

Using a combination of genetic analyses, environmental assessments and high-throughput DNA sequence classification of endogenous microbial populations in the mouth and gut, we examine how microbes may respond to and are responsible for environmental exposures, normal and disease states, as well as changes associated with aging. (colorado.edu)
In vitro overlay and immunoprecipitation analyses revealed preferential binding of the C-terminal sequences of CaV1.2 and ß2. (bvsalud.org)

Minipreparation of plasmid DNA is a rapid, small-scale isolation of plasmid DNA from bacteria. (wikipedia.org)

Unlike most protocols that use a 3-step lysis procedure, the QuickLyse Miniprep Kit combines enzymatic and osmotic processes to lyse bacterial cells in a single, 3-minute step. (qiagen.com)
Mechanical lysis involves the use of physical force, such as grinding or sonication, to break down bacterial cells and release the plasmid DNA. (wikipedia.org)

Although malaria in Yemen is caused mainly by Plasmodium falciparum and Plasmodium vivax , there are no sequence data available on the two species. (biomedcentral.com)
In conclusion, the homogeneity of the IgG-enriched species is postulated to represent a sequence that was strongly recognised by the humoral immune system at the time the sample was obtained. (biomedcentral.com)

If you don't have complete sequences send the information you have (maps, partial sequences etc. (customdnaconstructs.com)

From the 86 positive blood samples, 13 Plasmodium falciparum and 4 Plasmodium vivax were selected and underwent cloning and, subsequently, sequencing and the sequences were subjected to phylogenetic analysis using the neighbor-joining and maximum parsimony methods. (biomedcentral.com)
Following sequence analysis it was evident that the antibody depleted fraction was significantly more heterogeneous than the antibody enriched fraction, revealing a unique quasispecies profile. (biomedcentral.com)

Isolated plasmid DNA has OD 260 /280 ratios of 1.7-1.9, indicating that the quality and purity is suitable for all sensitive applications. (qiagen.com)
First, a suitable shRNA sequence must be identi﫿ed. (berterolab.com)

In addition, the provided wash plate also significantly reduces plasmid DNA cross-contamination between wells relative to other popular high-throughput plasmid prep kits. (zymoresearch.eu)

Viral RNA has been detected from skin lesions with the polymerase chain reaction (PCR) method in Sweden ( 15 ), but no genetic sequence is available. (cdc.gov)
Please review the details of this in the specific Sequencing Reaction section below. (dnaseq.co.uk)

Plasmid DNA is further purified by adsorption onto silica in the presence of high salts. (sigmaaldrich.com)
Presence or repetitive or high GC sequences, cloning involving large sequences or multiple changes may also increase the cost. (customdnaconstructs.com)

The QuickLyse Miniprep Kit is intended for molecular biology applications. (qiagen.com)

For the first time, minor differences within the sequence motif known as the VRN-box of VRN1 were correlated with wheat growth habit. (biomedcentral.com)

The sequencing results were rather promising except for the Hya promoter. (igem.org)
Because the sequencing results showed that the plasmid I sent did not contain the hya promoter I picked another colony that I incubated and from which I then isolated the plasmid again by a MiniPrep. (igem.org)

Plasmid-Safe DNAse and 10x PlasmidSafe Buffer (Lucigen, cat. (researchsquare.com)

The choice of which method to use will depend on the amount of plasmid DNA required, as well as the specific application for which it will be used. (wikipedia.org)
The purpose of this study was to generate oat expressed sequence tag (EST) information, develop a bioinformatics pipeline for SNP discovery, and establish a method for rapid, cost-effective, and straightforward genotyping of SNP markers in complex polyploid genomes such as oat. (biomedcentral.com)

A document with the complete plasmid sequence and confirmatory sequencing data of modified areas is emailed along with raw ABI sequencing files. (customdnaconstructs.com)

In addition, the wash regimen has been optimized to ensure the plasmid DNA is free of endotoxins, salt, protein, and RNA. (zymoresearch.eu)

A purified plasmid can be used for many standard applications, such as sequencing and transfections into cells. (wikipedia.org)
In addition, heat shocking the cells at 42°C creates a temperature gradient between the inside of the cell and its external environment, further assisting in sweeping the plasmids into the cells. (quartzy.com)
After you add the plasmid DNA to the cells, mix by gently flicking the tube 2-3 times. (quartzy.com)
We have made our tools and derivative intersectional mouse and ES cell lines openly available for non-commercial use through publicly curated repositories for plasmid DNA, ES cells, and transgenic mouse lines. (gointerlabs.com)

Anatomy2

Organisms25

Diseases4

Chemicals and Drugs41

Analytical, Diagnostic and Therapeutic Techniques and Equipment19

Phenomena and Processes69

Information Science4

Depending on the plasmid copy2

Purify plasmid4

Genome5

Purification14

Digestion3

Coli5

Primers6

Endotoxin-free3

Competent cells2

Clones2

Transfection2

Minipreps2

Restriction analysis1

Spin Column2

Buffers1

Genomic2

Gene6

Vector5

Klebsiella1

Amplification1

Yield1

Sufficient2

Constructs1

Alkaline1

RRNA1

Genes5

Assays1

Reactions2

Elution1

Nucleic1

Contigs1

Yields1

Primer6

Analyses2

Isolation1

Bacterial cells2

Species2

Partial1

Analysis2

Suitable2

Significantly1

Reaction2

High2

Molecular biology1

Motif1

Promoter2

Buffer1

Method2

Data1

Protein1

Cells4