Plasmid rnases. Medical search. Frequent questions

The inclusion of a good SOP (standard operating procedures) and the use of certified RNase-free reagents and equipment especially pipette tips contribute to removal of RNase. (severnbiotech.com)

Out of the seven tested recombinant human RNases, we have identified two members, RNase3 and RNase6, which were highly effective against Mycobacterium aurum extra- and intracellularly and induced an autophagy process. (frontiersin.org)
The recombinant plasmid D-pET32a-CP-His-FluA/B was transformed to BL21 with induction by IPTG. (flutrackers.com)
The set of tools under development at Blue Heron Bio will block functional protein expression and allow the propagation of recombinant plasmids using standard, high-copy vectors, potentially replacing the need for special growth conditions, low-copy vectors, special bacterial strains and other non-standard techniques. (technologynetworks.com)

In this process, the modified alkaline lysis method and RNase treatment are used for creating cleared lysate with minimal genomic DNA and RNA contaminants. (primelabmed.com)
It can be accessed online at www.helmholtz-hiri.de/en/datasets/enterococcus through an instance of the genomic viewer JBrowse. (frontiersin.org)
Analysis of plasmid, cosmid or genomic DNA in combination with various methods, e.g. (openwetware.org)
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)
RNase A is most used to remove RNA during the preparation of plasmid DNA or genomic DNA. (yeasenbiotech.com)

Qiagen) is often the major source of RNase contamination. (neb.com)
QIAGEN Ribonuclease A (RNase A) is endonuclease-free and quality-controlled for use in plasmid purification procedures for digestion of RNA. (qiagen.com)
This ready-to-use solution has the same specifications as the RNase supplied in all QIAGEN plasmid DNA purification kits. (qiagen.com)
EconoSpin® Plasmid Plus Giga Spin Column is compatible with Qiagen Plasmid Plus Giga Kit, Cat. (epochlifescience.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 RNase-Free DNase Set is guaranteed RNase-free, quality-controlled, and optimized for use with RNeasy procedures and with QIAamp RNA Blood Mini procedures. (bertec.com.tw)
The QIAGEN RNase-Free DNase Set is delivered as a stable, lyophilized enzyme. (bertec.com.tw)

Another timing issue caused by plasmids is that Cas enzyme and gRNA can remain active in the cells for prolonged periods. (idtdna.com)
This Sterile Highly Purified Water product is DEPC treated and manufactured to eliminate RNase enzyme activity. (severnbiotech.com)

Crude plasmid pellet containing pUC119 plasmid DNA was ethanol precipitated, suspended in TE buffer (lane 1), treated with RNase (lanes 2 and 5), and extracted with RapidClean once (lane 3) or twice (lane 4), or extracted with phenol-chloroform once (lane 6) or twice (lane 7). (gen.tr)

RNase contamination is not uncommon and can be very frustrating. (severnbiotech.com)
The techniques that are sources of contamination range from procedures such as plasmid preps to nuclease protection assays. (severnbiotech.com)
Other sources of RNase contamination can be bacterial cultures grown for cloning, or products, such as restriction enzymes. (severnbiotech.com)
Airborne or surface and solution contamination can also be a problem and the laboratory water supply is an obvious environmental source of RNase. (severnbiotech.com)

An aliquot of the solution must be diluted into Buffer P1 to the appropriate concentration as described in the relevant plasmid purification kit handbook. (qiagen.com)
dH 2 O, DNA, Restriction Endonuclease (10U/µL) and its appropriate (10x) Buffer, incubator (i.e. a water bath) @ 37°C, ice, micro-centrifuge tube(s) (MCT) Optionally: Ribonuclease A (RNase), Alkaline Phosphatase (ALP), incubator @ 65°C. (openwetware.org)
4. Add 5 μl TE buffer, place in a 42˚C water bath, and allow plasmids in the paper to elute for 20 minutes. (igem.org)
add 1 ml of PM1 Buffer to a RNase A tube, vortex the tube to dissolve the RNase A completely. (favorgen.com)
Transfer the total RNase A mixture back to the PM1 bottle, mix well by vortexing and store the PM1 buffer at 4 °C. (favorgen.com)

The RNase-Free DNase Set provides efficient on-column digestion of DNA during RNA purification from cells and tissues using RNeasy Kits and the QIAamp RNA Blood Mini Kit. (bertec.com.tw)
The RNase-Free DNase Set provides 1500 Kunitz units. (bertec.com.tw)

The purified plasmid (1 µg) is used in EcoRI digestion, and checked by electrophoresis. (primelabmed.com)

CRISPR plasmids expressing Cas enzymes and guide RNAs have been used for several years, but these plasmids have several limitations, including cytotoxicity and unpredictability of expression. (idtdna.com)

We further developed improved methods for circular RNA identification and purification using RNase R by identifying a way to better remove linear RNAs containing G-quadruplexes or structured 3' ends. (bcm.edu)

By revealing the fundamental mechanisms by which circular RNAs are generated, we have developed plasmid and viral-based methods for ectopically expressing circular RNAs. (bcm.edu)
CRISPR-Cas systems are a family of adaptive immune systems that use small CRISPR RNAs (crRNAs) and CRISPR-associated (Cas) nucleases to protect prokaryotes from invading plasmids and viruses (i.e., phages). (illinois.edu)

The Maxi Plasmid Kits use pre-packed ion-exchange resin columns to purify plasmid or cosmid DNA from bacterial cultures. (primelabmed.com)

MultiShot StripWell BL21 Star (DE3) Competent Cells offer enhanced mRNA stability due to a mutation in the RNaseE gene ( rne 131) that reduces levels of endogenous RNases and mRNA degradation, thereby increasing the stability of mRNA transcripts and increasing protein yield. (thermofisher.com)
Warning: mRNA sequence when devoid of trailing 'A's is still not a substring of the plasmid sequence. (iresite.org)
Plasmid containing full length 5' UTR of mouse Utrn mRNA (from nt -506 to nt -1 of the original sequence) and beta-galactosidase coding region. (iresite.org)

RNA synthesized from this kit is suitable for many applications including RNA structure and function studies, ribozyme biochemistry, probes for RNase protection or gel shift assays, hybridization-based blots, anti-sense RNA or RNAi experiments, microarray analysis, microinjection, sgRNA synthesis and in vitro translation studies. (neb.com)

The purified plasmid DNA is suitable for transfection, sequencing reactions, PCR, and in-vitro transcription. (primelabmed.com)
Some cell types cannot tolerate plasmid transfection, so one important consideration is direct cytotoxicity due to plasmid transfection itself. (idtdna.com)
Indeed, research shows that transfecting some cell types with any plasmid may cause cell death [ 1 ], while some transfection reagents (e.g., lipids) used for plasmid transfection are themselves toxic to cells [ 2 ]. (idtdna.com)

Once the target DNA has been successfully ligated into the plasmid vector, the plasmid must be transferred into the host cell for replication and cloning. (igem.org)

When CRISPR genome editing was introduced in 2009, plasmids were generously made available by the labs where they were developed so that other researchers around the world could use them to express Cas proteins in CRISPR experiments. (idtdna.com)
Importantly, it has also been shown that plasmid transfections, specifically in CRISPR experiments, can cause cytotoxicity in embryonic stem cells [ 3 ], which could be of concern to many researchers using this cell type. (idtdna.com)
Even with cells that can tolerate plasmid transfections, there are other problems associated with the use of plasmids for CRISPR editing, such as complications in the timing of CRISPR experiments. (idtdna.com)
The plasmid was used as the control for experiments in article The Utrophin A 5'-Untranslated Region Confers Internal Ribosome Entry Site-mediated Translational Control during Regeneration of Skeletal Muscle Fibers (Miura et al. (iresite.org)
I've successfully used the Zymoprep kit, but only for smaller plasmids in 2-hybrid experiments. (stackexchange.com)
In addition, this product can also be used in molecular biology experiments such as RNase protection analysis and RNA sequence analysis. (yeasenbiotech.com)

Armored RNA is a complex of MS2 bacteriophage coat protein and RNA produced in Escherichia coli by the induction of an expression plasmid that encodes the bacteriophage sequence consisting of the maturase, the coat protein, the pac site, and an exogenous RNA sequence. (politicalespionage.com)

Consider ribonucleoprotein (RNP) instead of CRISPR plasmids! (idtdna.com)
Ribonucleoprotein (RNP)-mediated CRISPR genome editing is more effective and avoids the problems associated with using plasmids. (idtdna.com)
Comparison of events after introducing CRISPR plasmid vs. ribonucleoprotein (RNP) into cells. (idtdna.com)
The ribonucleoprotein (RNP) form of archaeal RNase P comprises one catalytic RNA and five protein cofactors. (bvsalud.org)

Here, we identify RNase R as a third 'housekeeping' nuclease critical for immunity. (illinois.edu)
We show that RNase R works in concert with PNPase to complete crRNA maturation and identify specific interactions with Csm5, a member of the type III effector complex, which facilitate nuclease recruitment/stimulation. (illinois.edu)

Here we show, using both small interfering RNA (siRNA) particles and the plasmid based RNAI delivery system, pSUPER-RNAI, that decreasing endogenous hypothalamic AGRP levels in the central nervous system leads to significant physiological responses including an increase in metabolic rate and a decrease in body weight. (biomedcentral.com)

Using a gravity-flow procedure, the plasmid DNA in crude lysate has been bound to the column. (primelabmed.com)

While â ¼250-500 mM Mg2+ renders the archaeal RPR active without RNase P proteins (RPPs), addition of all RPPs lowers the Mg2+ requirement to â ¼10-20 mM and improves the rate and fidelity of cleavage. (bvsalud.org)

We strongly recommend adding 20 units Murine RNase Inhibitor ( NEB #M0314 ) in each reaction. (neb.com)
We measured on Northern gels the migration of reporter mRNAs isolated from cells transfected only with reporter plasmid or co-transfected with an active mutant of MAPK kinase-6, and treated either with or without the p38 MAPK inhibitor SB 203580. (ox.ac.uk)

Contribution of the enzymatic activity was discarded by site-directed mutagenesis at the RNase catalytic site. (frontiersin.org)
To catalyze Mg2+-dependent cleavage of the 5' leader from pre-tRNAs, the catalytic (C) and specificity (S) domains of the RNase P RNA (RPR) cooperate to recognize different parts of the pre-tRNA. (bvsalud.org)

If CRISPR-Cas plasmids were to be used in humans, these plasmids could pose hazards by integrating a Cas gene into the genome. (idtdna.com)
A 3V armored L-RNA of 2,248 bases containing six gene fragments-hepatitis C virus, severe acute respiratory syndrome coronavirus (SARS-CoV1, SARS-CoV2, and SARS-CoV3), avian influenza virus matrix gene (M300), and H5N1 avian influenza virus (HA300)-was successfully expressed by the two-plasmid coexpression system and was demonstrated to have all of the characteristics of armored RNA. (politicalespionage.com)

The total binding capacity of Plasmid Plus Giga column is 10 mg of plasmid DNA. (epochlifescience.com)

The FinO protein is essential for effective repression, acting by binding to FinP and protecting it from RNase E degradation. (wikipedia.org)
Armored L-RNA is a complex of MS2 bacteriophage coat protein and RNA produced in Escherichia coli by the induction of a two-plasmid coexpression system in which the coat protein and maturase are expressed from one plasmid and the target RNA sequence with modified MS2 stem-loop (pac site) is transcribed from another plasmid. (politicalespionage.com)

These plasmids have been cited in numerous publications and have helped to advance understanding and applications of CRISPR-Cas genome editing. (idtdna.com)
Meanwhile, however, many discoveries have been made about the best approaches to CRISPR genome editing-and it turns out that CRISPR plasmids are often not the most efficient approach to use, due to undesired effects such as cytotoxicity and unpredictability, both of which are discussed below. (idtdna.com)
Transfecting a plasmid creates a waiting time before the CRISPR genome editing can begin. (idtdna.com)

More than 150 labs have requested our circular RNA expression plasmids, highlighting our wide impact on this growing field. (bcm.edu)

I have to isolate a large plasmid from yeast and transform it in E. coli. (stackexchange.com)

RNase-resistant, noninfectious virus-like particles containing exogenous RNA sequences (armored RNA) are good candidates as RNA controls and standards in RNA virus detection. (politicalespionage.com)

3314-What is the concentration of RNase A sold separately? (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)

RNase A (17500 U), catalog number 19101, is 100mg/ml. (qiagen.com)
RNase A Solution (650 µl), catalog number 158922, and RNase A Solution (5 ml), catalog number 158924, are both 4mg/ml. (qiagen.com)

Template DNA, particularly plasmid DNA prepared by mini-prep (e.g. (neb.com)

For instance, only a few molecules of specific proteases, RNAses or ion channels can severely disrupt the growth or survival of a bacterial cell. (technologynetworks.com)

Reactions with 1 μg linearized plasmid DNA templates were incubated at 37°C in a PCR machine according to the manufacturer's recommendations. (neb.com)

Previously, we showed that the type III-A CRISPR-Cas system in Staphylococcus epidermidis employs two degradosome-associated nucleases, PNPase and RNase J2, to promote crRNA maturation and eliminate invading nucleic acids (Chou-Zheng and Hatoum-Aslan, 2019). (illinois.edu)

In contrast, RNAI can be used to study long-term phenotypes (including body weight and memory functions) due to the development of a DNA plasmid-based system to deliver short double-stranded RNA sequences in vivo [ 8 ]. (biomedcentral.com)

In conjugating bacteria the FinOP system regulates the transfer of F-like plasmids. (wikipedia.org)

The term "competent" is to describe a cell state in which there exist gaps or openings in the cell wall which will allow the plasmid containing the target genes to enter into the cell. (igem.org)

RNase A is the most active in cleaving single-stranded RNA and is active in a variety of reaction conditions: at low salt concentrations (0 to 100 mM NaCl), it can be used to cleave single-stranded RNA, double-stranded RNA, and RNA strands in RNA-DNA hybrids. (yeasenbiotech.com)

Differences in migration were shown by RNase H mapping with oligo(dT) to be due to poly(A) shortening. (ox.ac.uk)

RNase A is an endoribonuclease that specifically degrades single-stranded RNA at C and U residues. (yeasenbiotech.com)
Specifically, the cleavage recognizes the phosphodiester bond formed by the 5'-ribose of a nucleotide and the phosphate group on the 3'-ribose of the adjacent pyrimidine nucleotide, so that the 2', 3' - Cyclic phosphates are hydrolyzed to the corresponding 3'-nucleoside phosphates (eg, pG-pG-pC-pA-pG is cleaved by RNase A to generate pG-pG-pCp and A-PG). (yeasenbiotech.com)
while at high salt concentrations (≥0.3 M), RNase A can specifically cleave single-stranded RNA. (yeasenbiotech.com)

7. Add 5 μl RNase A (10 mg/ml) and incubate at 68C for 10 minutes. (igem.org)

The traJ 5' UTR is a cis acting RNA element which is involved in regulating plasmid transfer in bacteria. (wikipedia.org)

The addition of RNase might be considered if a small DNA fragment is expected that the RNA may otherwise obscure. (openwetware.org)

In this work, we have tested the human canonical members of the RNase family using a spot-culture growth inhibition assay based mycobacteria-infected macrophage model for evaluating their anti-tubercular properties. (frontiersin.org)

Anatomy8

Organisms39

Diseases3

Chemicals and Drugs86

Analytical, Diagnostic and Therapeutic Techniques and Equipment35

Phenomena and Processes93

Technology, Industry, Agriculture2

Information Science4

Health Care4

Certified RNase-free1

Recombinant3

Genomic5

Qiagen7

Enzyme2

Ethanol1

Contamination4

Buffer5

DNase2

Digestion1

Enzymes1

Purification1

RNAs2

Cosmid1

MRNA3

Assays1

Transfection3

Vector1

Experiments6

Encodes1

Ribonucleoprotein4

Nuclease2

Endogenous1

Crude1

Proteins1

Inhibitor2

Catalytic2

Gene2

Binding Capacity1

Protein2

Genome3

Expression1

Coli1

Particles1

Concentration1

Suitable1

Catalog2

Template DNA1

Molecules1

Reactions1

Eliminate1

Sequences1

System1

Target1

Active1

Shown1

Specifically3

Minutes1

Transfer1

Addition1

Family1