Buffer miniprep plasmids. Medical search. Frequent questions

Bacterial cell resuspension, lysis, and DNA binding are achieved in a single buffer, enabling fast DNA purification with minimal hands-on time. (qiagen.com)
Bacterial cells are resuspended and lysed and DNA is bound in a single buffer, enabling fast DNA purification with minimal hands-on time. (qiagen.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)
Compatible on a microcentrifuge or vacuum manifold for the purification of plasmid DNA. (zymoresearch.com)
Features polypropylene construction and contains a unique silica-based matrix that allows purification of up to 100 µg of plasmid DNA in ≥ 25 µl eluate when used in combination with ZymoPURE™ Plasmid buffers. (zymoresearch.com)
however, Miniprep I uses isopropanol precipitation and Miniprep II utilizes a column for purification. (mbpinc.net)
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)
For spin-column purification of endotoxin-free BAC/PAC plasmid DNA (up to ~200 kb) for sequencing, PCR, restriction endonuclease digestion, etc. (kyongshin.com)
1989). Plasmids were purified with Wizard plus SV miniprep DNA purification system (Promega, Madison, WI) according to the supplier's instructions. (her2signaling.com)
For purification of up to 50 micro g high-quality plasmid DNA in 96-well format. (labsave.com)
For Purification of up to 50micro g High-quality Plasmid DNA in 96-well Format, Includes TurboFilter and Plasmid Plus 96 Plates, Buffers, Reagents, Flat-Bottom Blocks, S-Blocks, Elution Microtubes. (labsave.com)
The High-Speed Plasmid Mini Kit was designed for both plasmid and cosmid DNA purification. (brbls.in)
Various methods have been developed for plasmid DNA purification. (sigmaaldrich.com)
GenElute™ Direct mRNA Miniprep kit for smaller-scale purification. (sigmaaldrich.com)
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)

Isolated from a strain of E. coli that carries a plasmid which encodes the β-Agarase I gene. (neuromantics.net)
Up to 100 µg per preparation, depending on the plasmid copy number, culture growth conditions, and strain of E. coli utilized. (zymoresearch.eu)
Neutralization/Clearing Buffer is designed to be used with our Zyppy-96 Plasmid Miniprep kit, which use a pellet-free modified alkaline lysis method to isolate ultra-pure plasmid DNA from E. coli. (zymoresearch.com)
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)
Eluted plasmid DNA can be used directly for E. Coli transformation, PCR, and Southern blot analysis. (mbpinc.net)
Plasmids are almost always purified from liquid bacteria cultures, usually E. coli, which have been transformed and isolated. (wikipedia.org)
The ligated plasmid was then introduced to E.coli by bacterial transformation, where they up took it via their cell membranes. (in2scienceuk.org)
Transform an aliquot of E. coli with an intact α-peptide-containing-plasmid (pBluescript for instance). (sigmaaldrich.com)
Escherichia coli strains XLBlue (recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proA+B+lacIqZ∆M15 Tn10 (TetR)] (Stratagene, La Jolla, CA), and CC118 [araD139 Δ(ara leu) 7697 ΔlacX74 phoAΔ20 galE galK thi rpsE rpoB argE(Am) recA1] (Manoil & Beckwith, 1986) were utilized SCH727965 as hosts for plasmids. (her2signaling.com)
Presented here is a protocol for the preparation and buffer calibration of cell extracts from exonuclease V knockout strains of Escherichia coli BL21 Rosetta2 ( ΔrecBCD and ΔrecB ). (jove.com)
We present the preparation steps of cell lysate from Escherichia coli BL21 Rosetta2 ΔrecBCD strain by sonication lysis and buffer calibration for Mg-glutamate (Mg-glu) and K-glutamate (K-glu) specifically for linear DNA. (jove.com)
This method is able to achieve protein expression levels comparable to that from plasmid DNA in E. coli CFPS. (jove.com)

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)

The QIAGEN QuickLyse Miniprep Kit uses one-step lysis technology for ultrafast preparation of up to 15 µg plasmid DNA. (qiagen.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)
The antigen is extracted from the cell in an appropriate lysis buffer, and antibodies are added to the lysate to allow formation of the immune complex. (neuromantics.net)
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)
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)
After adding 200μL of resuspention buffer, 200μL lysis buffer and 350μL neutralization buffer the solution was centrifuged for 10min. (igem.org)
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)
Enzymatic lysis, also called Lysozyme lysis, involves the use of enzymes to digest the cell wall and release the plasmid DNA. (wikipedia.org)
Commonly used miniprep methods include alkaline lysis and spin-column based kits. (wikipedia.org)
Innovative colored buffers for rapid identification of complete bacterial cell lysis and neutralization steps. (kyongshin.com)
The Lysis Buffer contains an integrated pH indicator to easily control the optimal pH value for DNA binding. (empiricalbioscience.com)
2006). Plasmid DNA was isolated from cultures grown in LB broth by an alkaline lysis method and visualized following electrophoresis in 1% agarose gels in TAE buffer (Sambrook et al. (her2signaling.com)
The High-Speed Plasmid Mini Kit uses a modified alkaline lysis method and RNase treatment to obtain clear cell lysate with minimal genomic DNA/RNA contaminants. (brbls.in)
Cells containing basic part plasmid DNA were lysed with the lysis device. (igem.org)
Lefty and righty cells containing plasmid DNA were lysed with our lysis device. (igem.org)
Since the lysis device results in successful release of plasmid DNA and assembly works in NEB2 buffered lysate, digestion of plasmid DNA in the lysate should work under the appropriate conditions. (igem.org)
The alkaline lysis-based miniprep protocol typically yields 5-15 μg of plasmid DNA from 1.5ml overnight cultures. (integra-biosciences.com)

Time for some plasmid isolation to be carried out. (igem.org)
Minipreparation of plasmid DNA is a rapid, small-scale isolation of plasmid DNA from bacteria. (wikipedia.org)
Empirical Bioscience Plasmid Mini-Prep Kit is designed for isolation of high-purity plasmid or cosmid DNA from bacterial cells for subsequent amplification, sequencing, restriction digests or transformations. (empiricalbioscience.com)
Preparing plasmid DNA depends on many time-consuming steps during cloning and large amounts of DNA isolation. (jove.com)

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)
This plasmid kit will purify plasmid DNA/cosmid DNA from 1-5 ml of cultured bacterial cells using an efficient plasmid miniprep system. (brbls.in)
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)

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)

In the presence of chaotropic salt, plasmid DNA in the lysate binds to the glass fiber matrix of the plasmid spin column. (brbls.in)
Commercial restriction enzymes and ligase were added to the lysate along with plasmid DNA. (igem.org)

Dulbecco's phosphate-buffered saline (DPBS) (Gibco, cat. (researchsquare.com)
Adhesion to Caco-2 The adhesion assay was set up in 24-well polystyrene plates as described previously [29], with only one modification: 2 × 102 cells in PBS (Phosphate Buffered Saline, Sigma) were added to each well. (gsk-3inhibitors.com)
E15.5 embryos were recovered and their brains dissected and fixed with 4% paraformaldehyde in phosphate-buffered saline. (bioinbrief.com)

Kit contents: Qiagen QIAprep 96 Plus miniprep Kit, 4 x 96 preps, 1.25 to 5mL Culture Volume, 48 to 96 Sample/Run, 40 min. (labsave.com)
Plasmid DNA mini preps and restriction enzyme digests are 'staples' in a laboratory that works with DNA. (rice.edu)

Can I substitute it with a homemade solution or Wash Buffer from another kit? (zymoresearch.eu)
Contaminants are removed with a Wash Buffer (containing ethanol) and the purified plasmid DNA is eluted by a low salt Elution Buffer, TE or water. (brbls.in)

The QuickLyse Miniprep Kit is intended for molecular biology applications. (qiagen.com)
Plasmids purified from genomic DNA, proteins, ribosomes, and the bacterial cell wall are used in molecular biology research. (sigmaaldrich.com)
The ease with which this DNA can be isolated and manipulated accounts for the widespread use of plasmids in molecular biology for a variety of tasks (e.g., protein expression). (rice.edu)

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

LigaFast Rapid DNA Ligation System (contains T4 DNA ligase and 2x Rapid ligation buffer, Promega, cat. (researchsquare.com)

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)

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)
It enables the extraction of plasmid DNA up to 10 kb length and yields up to 20 ?g DNA per preparation. (empiricalbioscience.com)
Typical yields are 20-30 μg for high-copy number plasmid or 3-10 μg for low-copy number plasmid from 4 ml of cultured bacterial cells. (brbls.in)

Q1: What is the composition of the ZymoPURE™ Elution buffer? (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)

It involved a lot of spinning in a centrifuge and adding different buffers. (in2scienceuk.org)
8 row reagent reservoir containing the different buffers for the protocol. (integra-biosciences.com)

General plasmid ligation protocol. (neuromantics.net)

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)
This plasmid kit does not require DNA phenol extraction or alcohol precipitation and the entire procedure can be completed within 15 minutes. (brbls.in)

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)

NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. (neb.com)

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)

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)

Ampicillin was added to the agar, as the bacteria with the plasmid also had ampicillin resistance so this was used to select them. (in2scienceuk.org)
Plasmids are small circles (usually less than 15 kb) of double stranded DNA maintained in some bacteria because they confer an advantage to the cells such as resistance to an antibiotic. (rice.edu)
Although plasmids occur naturally, the ones used in research have been engineered to prevent natural transfer between bacteria to reduce the spread of antibiotic resistance. (rice.edu)

Component in the ZymoPURE Plasmid Miniprep kit (Cat. (zymoresearch.com)

A few buffer conditions suit nearly all the restriction enzymes but no single buffer allows activity of every enzyme. (rice.edu)
Suppliers of enzymes always provide a reaction buffer (10x concentrate) that is optimum for the enzyme. (rice.edu)
Since restriction enzymes can require different buffer conditions, some strategy must be used to do double digests . (rice.edu)
The preferred method is to simultaneously digest with both enzymes in a compatible buffer. (rice.edu)

Prepare some LB agar plates containing the appropriate antibiotic to select for your chosen plasmid. (sigmaaldrich.com)

Reliably recover plasmid DNA from yeast colonies, patches on plates, or as liquid cultures. (mbpinc.net)

The unique vacuum plate design provides zero buffer retention and low elution volumes. (zymoresearch.eu)
Unique column design enables zero buffer retention and low-volume (≥ 10 μl) elution. (kyongshin.com)

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

A purified plasmid can be used for many standard applications, such as sequencing and transfections into cells. (wikipedia.org)
These were washed twice with sterile PBS (10 mM phosphate buffer, 2.7 mM potassium chloride, 137 mM sodium chloride, pH 7.4, Sigma), and resuspended in RPMI 1640 supplemented with morpholinepropanesulfonic acid (MOPS) at 1 × 106 cells/ml. (gsk-3inhibitors.com)
The experiment was repeated with lysed cells that were centrifuged and re-suspended in Buffer NEB2. (igem.org)
Cells were re-suspended in NEB2 Buffer. (igem.org)

Jena Bioscience Gel Loading Buffers with DNA Stain are formulated to facilitate loading of DNA samples into the wells of agarose and polyacrylamide gels. (jenabioscience.com)

Lysozyme is usually added to the bacterial culture, followed by heating and/or shaking the culture to release the plasmid DNA. (wikipedia.org)

We then carried out a restriction digest using the restriction enzyme EcoRI which cut the plasmid at a specific place. (in2scienceuk.org)
By definition, a unit of restriction enzyme will completely cleave 1µg of Lambda DNA (or other substrate DNA) in one hour in the recommended buffer and temperature. (rice.edu)

Homemade protocol, no kit is used Please check the buffer section first. (neuromantics.net)

Amplification of DNA fragments of the chr3N and chr3C genes from pUCywrB_A plasmid was carried out by PCR with oligonucleotides designed to yield translational PhoA/LacZ fusions within hydrophilic loops of the Chr3N and Chr3C proteins, according to a topological model based on hydropathic profiles and secondary-structure prediction programs. (her2signaling.com)

Green Gel Loading buffer with DNA Stain is recommended for analyzing DNA fragments 100 - 2000 bp. (jenabioscience.com)

On the other hand, PCR amplification from a plasmid, or a synthesized DNA template, can be used to simply prepare CFPS templates within a few hours 2 , 3 . (jove.com)

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

The buffers contain tracking dyes as indicator for DNA fragment migration. (jenabioscience.com)

Can Taq DNA Polymerase be used in other buffers? (genscript.com)
Taq DNA polymerase can have 100% activity in the supplied GenScript Taq Buffer. (genscript.com)
Additionally, the Taq DNA polymerase can also work well in other buffers. (genscript.com)

In extreme cases the DNA can be precipitated after one digest and dissolved in the second digest buffer. (rice.edu)

All these strategies cost extra time and resources and are yet to obtain near-plasmid levels of protein expression. (jove.com)

The eluted high-quality plasmid DNA is ready to use for a variety of down-stream applications. (empiricalbioscience.com)

The loading buffers contain SYBR® Green DNA Stain a fluorescent DNA intercalator dye specially developed for DNA analysis applications. (jenabioscience.com)

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

An alternative method is to digest with the 'low salt' enzyme then add more buffer and the 'high salt' enzyme to complete the digest. (rice.edu)

Anatomy3

Organisms30

Diseases4

Chemicals and Drugs53

Analytical, Diagnostic and Therapeutic Techniques and Equipment27

Phenomena and Processes80

Information Science4

Health Care2

Purification25

Coli12

Depending on the plasmid copy1

Lysis17

Isolation4

Purify4

Isolate4

Lysate2

Phosphate Buffer3

Preps2

Wash Buffer2

Molecular biology3

Vacuum manifold1

Promega1

Vectors1

Yields4

Elution Buffer2

Different buffers2

Ligation1

Extraction2

Genomic1

20211

Minipreps1

Vector1

Bacteria3

ZymoPURE1

Enzymes4

Antibiotic1

Cultures1

Zero buffer retention2

Strain1

Cells4

Agarose1

Bacterial culture1

Restriction enzyme2

Homemade1

Proteins1

20001

Amplification1

Transfection1

Fragment1

Polymerase3

Digest buffer1

Protein expression1

High-quality1

SYBR1

Typical1

Enzyme1