Large miniprep plasmids. Medical search. Frequent questions

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)
Various methods have been developed for plasmid DNA purification. (sigmaaldrich.com)
GenElute™ Direct mRNA Miniprep kit for smaller-scale purification. (sigmaaldrich.com)
We use the Promega Wizard Plus SV Minipreps DNA Purification System A1460 Miniprep protocol . (igem.org)
rAAV "Miniprep" includes production in one T-150 flask (150 cm 2 ) with partial purification by ultrafiltration, and quantitation by Free ITR qPCR. (ohsu.edu)
however, Miniprep I uses isopropanol precipitation and Miniprep II utilizes a column for purification. (mbpinc.net)
QIAfilter Plasmid Kits provide anion-exchange-based plasmid purification with bacterial lysate clearing by filtration. (qiagen.com)
The QIAfilter Plasmid Kits combines QIAfilter Cartridges for rapid clearing of bacterial lysates by filtration instead of centrifugation, with proven QIAGEN-tips containing anion-exchange resin for efficient plasmid purification. (qiagen.com)
QIAfilter Cartridges completely remove SDS precipitates and clear bacterial lysates in a fraction of the time needed for centrifugation, reducing plasmid-purification time by up to 1 hour. (qiagen.com)
The entire QIAGEN plasmid purification system avoids the use of toxic substances such as phenol, chloroform, ethidium bromide, and CsCl, minimizing hazard both to the user and the environment. (qiagen.com)
Genomic and plasmid DNA has to be removed during protein purification to produce DNA free Taq polymerase. (sydlabs.com)

The normal number of copies of plasmid that may be found in a single cell is called the plasmid copy number, and is determined by how the replication initiation is regulated and the size of the molecule. (wikipedia.org)

Yields of up to 10 mg (giga), 2.5 mg (mega), 500 µg (maxi), and 100 µg (midi) transfection-grade, high-copy plasmid DNA are achieved from culture (culture volumes depend on plasmid copy number, size of insert, host strain, and culture medium). (qiagen.com)

What is causing my problem with very low yields when isolating a 42kb yeast plasmid? (stackexchange.com)
The D200 Zymoprep-96 Yeast Plasmid Miniprep kit provides all the necessary reagents for high-throughput plasmid isolation from S. cerevisiae, C. albicans, S. pombe and any fungi whose cell walls are susceptible to yeast lytic enzyme lysis. (mbpinc.net)
The Zymoprep-96 is a simple and efficient yeast plasmid miniprep kit that is based on the classic E. coli alkaline lysis method with our Zymolyase added in the first solution. (mbpinc.net)
Order the D200 Zymoprep-96 Yeast Plasmid Miniprep kit in USA or in Canada from MBP Inc. for best rates and quick delivery. (mbpinc.net)
Q1: What is the difference between Zymoprep Yeast Plasmid Miniprep I and Miniprep II? (mbpinc.net)

High Copy Number = 100s of copies of plasmid per cell. (promegaconnections.com)

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)
Are there any other protocols that might be more suitable for a large plasmid and don't use too expensive materials? (stackexchange.com)
It might be worth trying the Qiagen yeast DNA protocols for the midi kit and see if you can isolate your large plasmid? (stackexchange.com)

The QIAGEN QuickLyse Miniprep Kit uses one-step lysis technology for ultrafast preparation of up to 15 µg plasmid DNA. (qiagen.com)
They use an expensive Qiagen large-construct column at the end, in order to purify and visualize the yeast extract on a gel, but I skip that step because the Qiagen column cost $40 per yeast clone which doesn't scale if we do many of these. (stackexchange.com)
The standard Qiagen midi columns are capable of capturing large constructs. (stackexchange.com)
The QIAGEN Plasmid Mega Kit (cat. (qiagen.com)
no. 12181) and the QIAGEN Plasmid Giga Kit (cat. (qiagen.com)
Visit the QIAGEN plasmid kits web page to learn more. (qiagen.com)
no. 19781) with the QIAGEN Plasmid Mega Kit (cat. (qiagen.com)
Prepacked QIAGEN-tips operate by gravity flow and never run dry, minimizing the hands-on time required for plasmid preparation. (qiagen.com)

Up to 100 µg per preparation, depending on the plasmid copy number, culture growth conditions, and strain of E. coli utilized. (zymoresearch.eu)
In the large-scale screen method, the partial colonies of E. coli bacteria can be directly transfered from the transformation plates into the solutions of the kits and immediately run on an agarose gel electrophoresis. (biochain.com)
I have to isolate a large plasmid from yeast and transform it in E. coli. (stackexchange.com)
This has worked wonders for me when I was purifying large BACmids from E. coli - and gets the cost down to $10/sample (it does require additional buffer, but that's a minor expense). (stackexchange.com)
Isolated from a strain of E. coli that carries a plasmid which encodes the β-Agarase I gene. (neuromantics.net)
Transform an aliquot of E. coli with an intact α-peptide-containing-plasmid (pBluescript for instance). (sigmaaldrich.com)
An impediment to the successful production of large quantities of natively folded proteins in Escherichia coli is the tendency of many proteins to become insoluble when overexpressed. (iucr.org)
Eluted plasmid DNA can be used directly for E. Coli transformation, PCR, and Southern blot analysis. (mbpinc.net)
This method is able to achieve protein expression levels comparable to that from plasmid DNA in E. coli CFPS. (jove.com)

Artificial plasmids are widely used as vectors in molecular cloning, serving to drive the replication of recombinant DNA sequences within host organisms. (wikipedia.org)
For our project, these insertion vectors will be low-copy BioBrick plasmids containing the integration site of a given phage, flanked by chosen restriction sites. (igem.org)
Our protein expression vectors are medium copy number plasmids and can be amplified using any standard miniprep or midi/maxi prep kits. (abmgood.com)

Q5: Can the ZymoPURE kits be used to isolate large plasmid constructs (BAC/PAC)? (zymoresearch.eu)
Works well with low-copy and hard-to-isolate plasmids. (mbpinc.net)
The system is ideal for low-copy numbers and hard-to-isolate plasmids. (mbpinc.net)
Q3: Can this kit be used to isolate linear plasmid DNA? (mbpinc.net)

The QuickLyse Miniprep Kit yields sufficient plasmid DNA for any downstream application, including automated sequencing. (qiagen.com)
High yields of high-quality plasmid DNA. (qiagen.com)
Specific yields for our plasmid DNA preparation services are offered anywhere from micrograms to milligrams in scale. (novoprolabs.com)

Preparing plasmid DNA depends on many time-consuming steps during cloning and large amounts of DNA isolation. (jove.com)

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)
So their cell lysis enzyme/buffer system works well, but I'd guess that your 42kb plasmid is precipitating out with the genomic DNA. (stackexchange.com)
QIAfilter Cartridges , provided in QIAfilter, HiSpeed, and EndoFree Plasmid Kits, are special filter units designed to replace centrifugation following alkaline lysis of bacterial cells. (qiagen.com)

The purified sample can be concentrated in volumes as low as 25 μL, in contrast to column-based kits which often necessitate larger elution volumes. (galenvs.com)
Most plasmid prep systems can process a range of volumes. (promegaconnections.com)

Plasmids purified from genomic DNA, proteins, ribosomes, and the bacterial cell wall are used in molecular biology research. (sigmaaldrich.com)

General plasmid ligation protocol. (neuromantics.net)

The term plasmid was introduced in 1952 by the American molecular biologist Joshua Lederberg to refer to "any extrachromosomal hereditary determinant. (wikipedia.org)
The QuickLyse Miniprep Kit is intended for molecular biology applications. (qiagen.com)
QIAfilter Plasmid Kits are intended for molecular biology applications. (qiagen.com)
Plasmid DNA preparation is one of the fundamental techniques of molecular biology research. (promegaconnections.com)

In the laboratory, plasmids may be introduced into a cell via transformation. (wikipedia.org)
BioChain's Express Cloning Checker Kits provide highly efficient methods of identifying recombinant colonies after transformation eliminating time-consuming plasmid preparation. (biochain.com)
Using linear DNA templates for CFPS will further enable the technology to reach its full potential, decreasing the experimental time by eliminating the steps of cloning, transformation, and plasmid extraction. (jove.com)

Reliably recover plasmid DNA from yeast colonies, patches on plates, or as liquid cultures. (mbpinc.net)
I then usually pick 2 colonies to grow up and miniprep. (visualizedlife.com)

Several tools and approaches have been developed to identify plasmid sequences in such assemblies, but require trade-off between sensitivity and specificity. (gointerlabs.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)
6). Detection of specific nucleic acid sequences by hybridization a. mini-prep of plasmid DNA b. (ntu.edu.tw)
This will require secondary DNA sequences including the phage genome integration site , some garbage DNA (flanked by essential restriction enzyme cut sites) and the low-copy number BioBrick plasmid . (igem.org)
Presence or repetitive or high GC sequences, cloning involving large sequences or multiple changes may also increase the cost. (customdnaconstructs.com)
Using a really basic U6-promoter guide-RNA expressing plasmid (like this ), design a pair of long primers that overlap in their 5′ ends with the guide-RNA sequence, but hybridize to sequences adjacent to the guide-RNA sequence but point in the opposite directions. (visualizedlife.com)

The results will be known within 1-2 hours instead of the 2 days required in a conventional method of labor-intensive plasmid DNA miniprep or colony hybridization. (biochain.com)
A blue coloured colony indicates that the α-peptide in the plasmid is intact (no insert) whereas a white colony indicates that the α-peptide is disrupted (insert present). (sigmaaldrich.com)
If you will be using 100-1,000 ml of culture, pick an isolated colony from a freshly streaked plate, inoculate 5-50 ml of LB medium [+ antibiotic(s)] and grow for 8 hours to overnight at 37 °C. Next, inoculate a larger volume of culture medium [+ antibiotic(s)] by diluting the starter culture 100- to 500-fold. (promegaconnections.com)
When I run Sanger, usually at least one colony contains plasmid DNA with the intended guide-RNA encoded. (visualizedlife.com)
If neither colony has the intended plasmid, I normally pick two more and screen those. (visualizedlife.com)

I've successfully used the Zymoprep kit, but only for smaller plasmids in 2-hybrid experiments. (stackexchange.com)
The Zymoprep-96 incorporates our Zymoprep II kit into a 96-well format resulting in a high-throughput method that recovers about 5 fold more plasmid DNA compared to Zymoprep I and eliminates the isopropanol precipitation step. (mbpinc.net)

You can find out what bacterial strain carries the plasmid by looking over your confirmation email-it will appear next to the plasmid name. (dictybase.org)
Then transform the plasmid into the host bacterial strain and expose the bacteria to the phage. (igem.org)

Through phosphorothioate modification of primers, we realized onepot detection with high sensitivity using plasmids of SARS-CoV-2, HPV16 and HPV18. (gointerlabs.com)

Eluted Plasmid DNA is Transfection-Grade and Free of Cross-Contamination. (zymoresearch.eu)

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)
We report that the variable domains of recombinant HIV-1 Env immunogens activate a large number of B cell clones that give rise to many non-neutralizing antibodies, and that removing the variable domains from the immunogen reduces the number of activated B cell lineages and leads to the development of autologous neutralizing antibodies, a step towards bNAb-production. (plos.org)

Answer: It is always heard that scientists can put a Taq polymerase gene into an expression vector and produce large quantity of Taq enzyme in their labs with low cost. (sydlabs.com)
The replicon controls plasmid replication by bacterial enzyme complexes, controlling the plasmid copy numbers, and thus DNA yield. (promegaconnections.com)

In nature, plasmids often carry genes that benefit the survival of the organism and confer selective advantage such as antibiotic resistance. (wikipedia.org)
Prepare some LB agar plates containing the appropriate antibiotic to select for your chosen plasmid. (sigmaaldrich.com)
2. Information on antibiotic resistance of the plasmid. (novoprolabs.com)

The cleared lysate is then loaded onto the anion-exchange tip where plasmid DNA selectively binds under appropriate low-salt and pH conditions. (qiagen.com)

Plasmids are considered replicons, units of DNA capable of replicating autonomously within a suitable host. (wikipedia.org)
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)

A typical bacterial replicon may consist of a number of elements, such as the gene for plasmid-specific replication initiation protein (Rep), repeating units called iterons, DnaA boxes, and an adjacent AT-rich region. (wikipedia.org)
In addition, the wash regimen has been optimized to ensure the plasmid DNA is free of endotoxins, salt, protein, and RNA. (zymoresearch.eu)
Celetrix cell electroporation large-scale model SLT (catalog No. 11-0104) is a unique CHO electroporation machine for large scale protein transient production in CHO cells. (ushelf.com)
It's also nice in that if the change is made, even if the repair is an in-frame indel, the deletion will be so large that the protein should still be loss-of-function. (visualizedlife.com)
All these strategies cost extra time and resources and are yet to obtain near-plasmid levels of protein expression. (jove.com)

This host-to-host transfer of genetic material is one mechanism of horizontal gene transfer, and plasmids are considered part of the mobilome. (wikipedia.org)
Plasmids almost always carry at least one gene. (wikipedia.org)
A: Specify what gene or destination plasmid you need and indicate that you don't have source plasmids. (customdnaconstructs.com)
Lenti-SV40 will contain the whole SV40 gene, -SV40T, the large T Antigen only, and -SV40T&t the large and small T antigens only. (abmgood.com)

Lastly, lysogenic phage genomes are too large to be manipulated using normal BioBrick plasmids. (igem.org)
This will allow the phage to infect the bacterial cells as well as the integration site on the low-copy BioBrick plasmid. (igem.org)
Current state of the low copy BioBrick plasmid containing the full phage genome. (igem.org)

Perform 96 Plasmid Minipreps Simultaneously using a Vacuum Manifold or Centrifuge. (zymoresearch.eu)

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)

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)

3) "Purify" your intended plasmid. (visualizedlife.com)

For insert cloning please send the sequence of the insert, or, better, the complete sequence of the source plasmid for the insert. (customdnaconstructs.com)
You can also describe the required changes by simply sending the full sequence of the desired final plasmid. (customdnaconstructs.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)
1) Amplify plasmid, appending the guide-RNA sequence to the termini of the amplicon. (visualizedlife.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)

Production size can be anywhere from a small pilot stock to a large prep, with purity levels varying from crude lysate to highly purified stock. (ohsu.edu)

A: Plasmids are prepared as miniprep DNA, a small amount, sufficient for multiple transformations, is sent in 10ul TE. (customdnaconstructs.com)

The term's early usage included any bacterial genetic material that exists extrachromosomally for at least part of its replication cycle, but because that description includes bacterial viruses, the notion of plasmid was refined over time to comprise genetic elements that reproduce autonomously. (wikipedia.org)
In order for plasmids to replicate independently within a cell, they must possess a stretch of DNA that can act as an origin of replication. (wikipedia.org)
Smaller plasmids make use of the host replicative enzymes to make copies of themselves, while larger plasmids may carry genes specific for the replication of those plasmids. (wikipedia.org)

NovoPro plasmid DNA preparation services are accommodating to small research labs as well as large-scale manufacturing biotech and pharmaceutical companies. (novoprolabs.com)

Plasmids are generally circular, but examples of linear plasmids are also known. (wikipedia.org)
However, so far CFPS systems rely on circular plasmids that can limit the technology from reaching its full potential. (jove.com)

Routine PCR using genomic, viral and plasmid templates. (sydlabs.com)

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)

Using standard BioBrick assembly methods, ligate the integration site onto a low-copy BioBrick plasmid (Fig. 3). (igem.org)
Prepare a high copy BioBrick plasmid with the A1 and Unique site by PCRing these sites onto some garbage DNA and cutting/ligating the DNA onto the plasmid. (igem.org)
The next step is to cut at A1 and the Unique site and ligate this DNA onto the higher copy garbage (with cut sites) plasmid. (igem.org)
After culture conditions and bacterial host, the copy number is the next factor that influencse plasmid DNA yield. (promegaconnections.com)

Microvolume applications include plasmid, DNA and RNA work, and miniprep. (globalspec.com)

From this culture, miniprep the DNA - very carefully since it is now a 50 kilo base pair, or more, plasmid. (igem.org)
For example, the PureYield™ Miniprep System can process 600μl to 3 ml of culture. (promegaconnections.com)

Many of the genes carried by a plasmid are beneficial for the host cells, for example: enabling the host cell to survive in an environment that would otherwise be lethal or restrictive for growth. (wikipedia.org)
Some plasmids, however, have no observable effect on the phenotype of the host cell or its benefit to the host cells cannot be determined, and these plasmids are called cryptic plasmids. (wikipedia.org)
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)
No special competent cells required, as they are stable plasmids. (abmgood.com)
Instead, I'll describe a likely less optimal, but super cheap and easy method of co-transfecting Cas9 and guide-RNA expressing plasmids into cells to create knockouts. (visualizedlife.com)
To begin to define the specificity determinants of RGENs in human cells, we sought to perform a large-scale test in which we assessed the effects of systematically mismatching various positions within multiple gRNA/target DNA interfaces. (cdc.gov)

While chromosomes are large and contain all the essential genetic information for living under normal conditions, plasmids are usually very small and contain only additional genes that may be useful in certain situations or conditions. (wikipedia.org)
Plasmids play an important role in bacterial evolution and mediate horizontal transfer of genes including virulence and antimicrobial resistance genes. (gointerlabs.com)

Plasmids vary in size from 1 to over 400 kbp, and the number of identical plasmids in a single cell can range anywhere from one to thousands under some circumstances. (wikipedia.org)
Their size can range from very small mini-plasmids of less than 1-kilobase pairs (kbp) to very large megaplasmids of several megabase pairs (Mbp). (wikipedia.org)
What is the largest plasmid size that can be purified using the QuickLyse Miniprep Kit? (qiagen.com)
The QuickLyse Miniprep Kit can be used for plasmids up to 9 kb in size. (qiagen.com)

Q2: What is the typical plasmid yield? (mbpinc.net)

Anatomy2

Organisms25

Diseases4

Chemicals and Drugs41

Analytical, Diagnostic and Therapeutic Techniques and Equipment19

Phenomena and Processes69

Information Science4

Purification12

Called the plasmid copy1

High-copy plasmid1

Yeast plasmid5

Copies of plasmid1

Protocols3

QIAGEN8

Coli9

Vectors3

Isolate4

Yields3

Isolation1

Lysis3

Volumes2

Genomic DNA1

Ligation1

Molecular4

Transformation3

Colonies2

Sequences6

Colony5

Zymoprep2

Bacterial strain2

Primers1

Transfection1

Recombinant2

Enzyme2

Antibiotic3

Binds1

Suitable2

Protein5

Gene4

Phage3

Centrifuge1

Centrifugation1

Buffers1

Purify1

Sequence4

Prep kits1

Lysate1

Sufficient1

Replication3

Preparation1

Circular2

Viral1

Genome1

Copy4

Applications1

Culture2

Cells6

Genes2

Size4

Typical1