Mammalian cloning efficiency. Medical search. Frequent questions

High-efficiency full-length cDNA cloning. (nih.gov)
The SuperScript IV CellsDirect cDNA Synthesis Kit is designed to synthesize first-strand cDNA directly from mammalian cell lysates without first isolating the RNA. (thermofisher.com)
Discover the largest collection of ORF/cDNA expression clones from different species, including human, mouse, rat, and more with GeneMedi's Promise-ORF TM . (genemedi.net)
1995). This technique consists of the large-scale, single-pass, and incomplete sequencing of cDNA clones (around 500 bp), from a lot of libraries representing diverse tissues usually. (colinsbraincancer.com)
First, a full-length functional ZIKV cDNA clone was engineered as a bacterial artificial chromosome, with each reporter gene under the cap-independent translational control of a cardiovirus-derived internal ribosome entry site inserted downstream of the single open reading frame of the viral genome. (mdpi.com)

A T7 expression plasmid and the same plasmid containing a gene encoding a toxic mammalian protein were transformed into each host. (neb.com)
Small interfering RNAs (siRNAs) expressed from short hairpin RNAs (shRNAs) are a powerful way to mediate gene specific RNA interference (RNAi) in mammalian cells. (sigmaaldrich.com)
The neoplastic potential of transformed cells was determined via soft agar cloning and gene transfection analysis. (cdc.gov)
Get inducible gene expression for mammalian systems with SBI. (systembio.com)
Simply email us the sequence of your gene and we'll synthesize and clone it into any of our inducible vectors for little more than the cost of the empty vector. (systembio.com)
2） Stable expression of fluorescent gene or resistant gene ，benefit for subsequent experiment such as antibiotic screening and single-cell clone isolation. (ubigene.us)
expression Mammalian Cepko 8389 p212 pCMV-EGFP/RFP EGFP- dsRed gene switch plasmid Mammalian Green 22799 Ai9 Rosa26. (addgene.org)
A putative acylphosphatase gene (PhAcP) was cloned from the genomic DNA of Pyrococcus horikoshii and was expressed in Escherichia coli. (rcsb.org)
No. 632602) is a complete system for the cloning and expression of a target single guide RNA (sgRNA) for mammalian gene modification studies using CRISPR/Cas9 technology. (biofeng.com)
Serial analysis of gene expression (SAGE) is really a widely used way of large-scale transcriptome analysis in mammalian systems. (colinsbraincancer.com)
Based upon an equal result in the tests of all oils from the Citrus category, the results of the available gene mutation tests in mammalian cells with a citrus oil can be read across to the other oils. (europa.eu)
For both orange oil and lemon oil, the gene mutation study in mammalian cells was negative. (europa.eu)
A protein coded by a variant gene may be dysfunctional, which can affect the efficiency and safety of the substrate drug individually. (helsinki.fi)
For expression in the Sf9 cells, the aim of the study was to clone the SLCO1B1-gene into the pFastBac vector. (helsinki.fi)

Therapeutic monoclonal antibodies (MAbs) mostly are manufactured using bioengineered mammalian cells cultured in a bioreactor for two to three weeks. (bioprocessintl.com)

donor plasmid backbones pSL1180-HR-PUbECFP & pSL1180polyUBdsRED PDF 583.3 KB Zhang gRNA cloning CRISPR RNA. (addgene.org)

These ATG genes are highly conserved in mammalian cells, allowing to study their functions also in higher eukaryotes [ 1 - 4 ]. (hindawi.com)
In contrast, recombinant antibodies are monoclonal antibodies that are generated by cloning antibody genes into expression vectors and do not involve the use of hybridomas. (sigmaaldrich.com)
Use of a single vector carrying two separate expression cassettes provides higher efficiency and both genes are integrated at the same position and expressed independently. (sigmaaldrich.com)
For this study, newly developed scFv genes were cloned into different mammalian expression vectors and transferred into competent cells by chemical transformation. (itu.edu.tr)

Lentiviral vectors can be used to deliver shRNAs, thereby providing the ability to infect most mammalian cell types with high efficiency, regardless of proliferation state. (arizona.edu)
Lentiviral inducible shRNA vectors are readily available, but unfortunately the process of cloning, screening, and testing shRNAs can be time-consuming and expensive. (arizona.edu)
Then, the cloned vectors were determined by colony polymerase chain reaction ( PCR) studies. (itu.edu.tr)
Our collection of ORFs expression clones come in verified mammalian or viral vectors lentivirus, AAV, and adenovirus) and our Promise-ORF TM viral-ready expression vectors make it easy for you to improve viral vector packaging efficiency or directly transfect mammalian cells, resulting in stronger and easier expression detection. (genemedi.net)

Among the factors thought to contribute to the greater success in cloning cattle are the relatively late embryonic genome activation speciﬁc for this species [16 -18] and the optimization of reproductive technologies, such as in vitro embryo production and embryo transfer, brought about by the cattle industry [19]. (sibi.org)
Cloning and Expression System for sgRNAs The Guide-it CRISPR/Cas9 Systems are kits for the cloning and expression of target single guide RNAs (sgRNAs) for mammalian genome editing using CRISPR/Cas9 technology. (biofeng.com)
To facilitate mammalian genome engineering applications, we provide a web-based software tool to guide the selection and validation of target sequences as well as off-target analyses. (cdc.gov)

Our electroporation cuvettes are designed for use with bacterial, yeast, fungi, and mammalian cells. (thomassci.com)
Use for bacterial transformations, eukaryotic transfections, and the electroporation of mammalian cells and yeast. (thomassci.com)

High efficiency chemically competent E. coli cells suitable for T7 expression with enhanced reduction of basal expression. (neb.com)
Enhanced BL21 derivative, chemically competent E. coli cells suitable for high efficiency transformation and protein expression. (neb.com)
50 μl of competent cells were transformed with 100 pg of pUC19 control DNA following the provided High Efficiency Transformation Protocol except heat shock time varied from 0 to 80 seconds. (neb.com)
In addition, the lentiviral transduction particles are pseudotyped with an envelope G glycoprotein from vesicular stomatitis virus (VSV-G), allowing transduction of a wide variety of mammalian cells. (sigmaaldrich.com)
So far the reprogramming of somatic cells shows very low rates of efficiency (~0.0006-1%) that have not improved in the last two decades of continuous research. (sibi.org)
Cloning by nuclear transfer using mammalian somatic cells has enormous potential application. (biomedcentral.com)
Various strategies have been employed to modify donor cells and the nuclear transfer procedure in attempts to improve the efficiency of nuclear transfer. (biomedcentral.com)
Recombinant antibody production from most species consist of a heavy and a light chain and it is important to note that to accomplish maximum IgG production in mammalian cells, one must achieve a balanced expression of both heavy and light chains. (sigmaaldrich.com)
The CloneSelect® Imager FL is a high-throughput automated solution for imaging and analyzing mammalian cells. (moleculardevices.com)
3）Lentivirus has a wide range for host and can effectively infect all mammalian cells. (ubigene.us)
In this manner, two different approaches have been adopted in the thesis, the first is the production and control of the target scFv molecules in mammalian cells, and the second approach is to transfer the target scFv molecules to a different structure using the complementarity determining regions CDR-grafting method, ensuring their production in bacteria and controlling their activity. (itu.edu.tr)
This renders cells auxotrophic for the essential amino acid L-glutamine and, therefore, speeds up and simplifies the selection and scale-up of clones. (iptonline.com)
The included vector is used to simultaneously express Cas9, a target-specific sgRNA, and an exceptionally bright fluorescent protein (either ZsGreen1 or tdTomato) for monitoring transfection efficiency and/or for further enriching/isolating transfected cells by flow cytometry. (biofeng.com)
The vector in this system simultaneously expresses Cas9 nuclease, a target-specific sgRNA, and an exceptionally bright fluorescent protein for monitoring transfection efficiency and/or for further enriching/isolating transfected cells by flow cytometry (ZsGreen1 and tdTomato versions are available). (biofeng.com)
The kit also includes Stellar competent cells to ensure high efficiency transformation. (biofeng.com)
display libraries from electroporation-competent cells with high transformation efficiency. (thomassci.com)
These cells also offer the highest electroporation efficiency (Hee) phenotype, which confers increased resilience in harsh electroporation conditions. (thomassci.com)
Experiments with cynomolgus monkey iPSC-derived PGCLCs have allowed us to analyze germ lineage development across different mammalian species and identify the early amniotic membrane as the origin of germ cells in primates. (kyoto-u.ac.jp)
Human Embryonic Kidney cells (HEK) is a human derived mammalian cell-line that is widely used in the study of OATP1B1 transporter. (helsinki.fi)
His work has led to major advances in our understanding of mammalian cloning and embryonic stem cells. (wraltechwire.com)

The efﬁciency of cloning, deﬁned as the proportion of transferred embryos that result in viable offspring, is approximately 2 to 3% for all species. (sibi.org)
However, in cattle, average cloning efﬁciency is higher than in other species, ranging from 5 to 20% [10 -15]. (sibi.org)
However, somatic cloning has been inefficient in all species in which live clones have been produced. (biomedcentral.com)
Because cattle are a species widely used for nuclear transfer studies, and more laboratories have succeeded in cloning cattle than any other specie, this review will be focused on somatic cell cloning of cattle. (biomedcentral.com)
These antibodies can be cloned from any species using suitable oligonucleotide primers. (sigmaaldrich.com)
Several mutations in LHR have been identified in many mammalian species, leading to either constitutive activation or inactivation of the receptor. (preprints.org)

Cell cultures derived from mammalian and bacterial cell lines are the conventional production systems in bioprocessing. (bioprocessintl.com)

Subsequently, two ubiquitin-like conjugation systems, ATG5/ATG12 and ATG8, the mammalian homologues of which include LC3, GABARAP, and GATE-16, concert the formation of the double-membraned autophagosome [ 9 ]. (hindawi.com)
Our data suggest that, while PhAcP is less efficient than other mammalian homologues at 25 degrees C, the thermophilic enzyme is fully active at the optimal growth temperature (98 degrees C) of P. horikoshii. (rcsb.org)

This shRNA non-mammalian control was designed using our Turbo GFP sequence and may cause some knockdown of tGFP. (sigmaaldrich.com)
In these cell lines, MISSION shRNA clones permit rapid, cost efficient loss-of-function and genetic interaction screens. (sigmaaldrich.com)
When conducting experiments using MISSION ® shRNA clones, the proper controls should be a key element of your experimental design to allow for accurate interpretation of knockdown results. (sigmaaldrich.com)
Results: Notably, we emphasize the importance of loop sequence design and demonstrate that the addition of a single mismatch in the loop stem can greatly improve shRNA efficiency. (arizona.edu)
Conclusions: Our aim was twofold: first, to take a very useful shRNA vector and make it more amenable for molecular cloning and, secondly, to provide a streamlined protocol and rationale for cost-effective design, cloning, and screening of shRNAs. (arizona.edu)

Antisense transcription in the mammalian transcriptome. (nih.gov)

a pair of user-provided oligos corresponding to the target genomic sequence of interest are annealed to form a duplex, and the duplexed oligos are inserted into the pre-linearized vector using the included high-efficiency ligation mix. (biofeng.com)
The enzyme and buffer system are room temperature stable for 4 weeks and give superior PCR performance on complex templates such has mammalian genomic DNA. (witec.ch)

Pre-made anti-EZH2 Y641F monoclonal antibody(mab) is expressed by mammalian cell line as a benchmark antibody for cell culture, ELISA or other affinity binding assay or functional assay development, animal model development, PK/PD model development (Pharmacokinetics & Pharmacodynamic). (genemedi.net)
The anti-EZH2 Y641F mab is expressed and produced by mammalian cell line as a benchmark reference therapeutic antibody for biological drug discovery items including cell culture, assay development, animal model development, PK/PD model development (Pharmacokinetics & Pharmacodynamic) and mechanism of action (MOA) research. (genemedi.net)

True cloning performed by nuclear transfer from an adult and differentiated somatic cell to a previously enucleated egg (somatic cell nuclear transfer, SCNT), gives rise to a new cell, the nuclovulo (nucleus+ovum), distinct from the zygote because the sperm is not involved in its creation, while both can develop as embryos and give rise to offspring. (sibi.org)

An optimized lentiviral vector system for conditional RNAi and efficient cloning of microRNA embedded short hairpin RNA libraries. (arizona.edu)

recombination to maximize label diversity Mammalian Megason 13769 pCALNL- DsRed Cre dependent DsRed expression. (addgene.org)
Mammalian Expression DsRed2 563 582 24 Tetramer pCAG- DsRed2 - Mammalian Expression DsRed2 -N1 - Mammalian Expression. (addgene.org)
In the first approach, the target scFv protein expression in mammalian cell culture was investigated. (itu.edu.tr)
Commonly used cell lines include Chinese hamster ovary (CHO) mammalian cell expression systems, such as CHOK1. (iptonline.com)

Relative colony forming efficiencies (RCE) were used to determine glass fiber cytotoxicity. (cdc.gov)
The enhanced efficiency of PCRBIO HiFi Polymerase minimises PCR inhibition, from impure samples such as colony PCR and direct PCR. (witec.ch)

Do the cloning and transductions/transfections yourself or leave the cell line engineering to SBI. (systembio.com)

Currently, the efficiency for nuclear transfer is between 0-10%, i.e., 0-10 live births after transfer of 100 cloned embryos. (biomedcentral.com)

Comparison of the relative transformation efficiencies demonstrates that the T7 Express hosts provide the levels of control necessary for transformation of potentially toxic clones. (neb.com)
Biochemical reconstitution demonstrates that the mammalian complex possesses inherent E3 ubiquitin ligase activity, using Ube2H as its cognate E2. (elifesciences.org)

shRNAs that generate siRNAs intracellularly are expressed from amphotropic lentivirus viral particles, allowing screening in a wide range of mammalian cell lines. (sigmaaldrich.com)

GeneArt Gibson Assembly cloning is the optimal choice for building large and demanding constructs from multiple DNA fragments. (thermofisher.com)
The DNA sequences of the newly cloned scFv constructs were controlled by a capillary electrophoresis system. (itu.edu.tr)

Cas-Designer searches for targets that maximize knockout efficiency while having a a low probability. (addgene.org)
For all five tested targets, we observed a consistent increase in modification efficiency with increasing tracrRNA length. (cdc.gov)

They are ideal for mammalian and insect cell culture. (tritechresearch.com)

Hence, a monoclonal antibody is considered a collection of identical antibodies secreted by a single B cell clone with specificity for only one antigenic epitope. (sigmaaldrich.com)

Maximize efficiency, preserve cell viability, and streamline your workflow with DispenCell and CloneSelect Imager FL bundle. (moleculardevices.com)
Whatever your workflow needs, our comprehensive and diverse molecular solutions are designed for high performance, time savings, and reliability for your PCR and cloning success. (thermofisher.com)

Methods: First, we modified the Tet-pLKO-Puro vector to make it easy("EZ") for molecular cloning (EZ-Tet-pLKO-Puro). (arizona.edu)
We have also generated PGCLCs from human iPSCs, having compared the induction efficiencies for germ lineage between iPSC clones and found the molecular and signaling factors responsible for the different efficiencies. (kyoto-u.ac.jp)

Here, we identified all subunits of the mammalian GID/CTLH complex and provide a comprehensive map of its hierarchical organization and step-wise assembly. (elifesciences.org)

Twenty years have passed since Dolly the sheep was born by cloning (somatic cell nuclear transfer, SCNT) but the results of non-human mammalian cloning are very poor, and cause animal diseases and huge biological losses. (sibi.org)
Various strategies have been used to improve the efficiency of nuclear transfer, however, significant breakthroughs are yet to happen. (biomedcentral.com)
Somatic cell cloning (cloning or nuclear transfer) is a technique in which the nucleus (DNA) of a somatic cell is transferred into an enucleated metaphase-II oocyte for the generation of a new individual, genetically identical to the somatic cell donor (Figure 1 ). (biomedcentral.com)
Although the efficiency of nuclear transfer has been dramatically improved from the initial success rate of one live clone born from 277 embryo transfers [ 1 ], none of the aforementioned efforts abolished the common problems associated with nuclear transfer. (biomedcentral.com)

Therefore, we sought to refine a popular vector (Tet-pLKO-Puro) and streamline the cloning process with efficient protocols so that researchers can more efficiently utilize this powerful tool. (arizona.edu)

user clones guide sequence' rep_origin complement(147. (biofeng.com)

The cloning was not successful in this study although attempts were made for several approaches. (helsinki.fi)

4th, concatemers were partly digested with Nla III before cloning into vector (pZEro-1), improving cloning efficiency greatly. (colinsbraincancer.com)

For an extraordinary record of innovations in improving the energy efficiency and reducing the cost of gas liquefaction and separation. (wraltechwire.com)

dramatically improving cloning large plasmids and ligated DNA. (thomassci.com)

These developmental defects have been attributed to incomplete reprogramming of the somatic nuclei by the cloning process. (biomedcentral.com)

Methods and technical offerings in the biopharmaceutical contract development and manufacturing space have led to dramatic time reductions in CLD and further efficiencies in upstream and downstream processes. (iptonline.com)

In a meeting in Washington (3 December 2001) the researcher Tanja Dominko presented the results of monkey cloning (Macacus rhesus) when she worked at the Regional Center of Research in Primates of Beaverton, Oregon (USA). (sibi.org)
Developmental defects, including abnormalities in cloned fetuses and placentas, in addition to high rates of pregnancy loss and neonatal death have been encountered by every research team studying somatic cloning. (biomedcentral.com)
With Applied Biosystems and Invitrogen products for PCR and cloning, there is no wondering whether they will propel your research forward. (thermofisher.com)

Why Cloning in Non-Human Mammalians Fail? (sibi.org)

Anatomy7

Organisms2

Diseases1

Chemicals and Drugs5

Analytical, Diagnostic and Therapeutic Techniques and Equipment12

Phenomena and Processes10

Information Science3

Health Care2

CDNA5

Gene14

Bioengineered mammalian cells1

Plasmid1

Genes4

Vectors4

Genome3

Yeast2

Cells20

Species6

Bacterial1

Homologues2

ShRNA5

Transcriptome1

Genomic2

Assay2

Differentiated somatic1

RNAi1

Expression4

Colony2

Transfections1

Embryos1

Demonstrates2

Lentivirus1

Constructs2

Targets2

Insect1

Antibody1

Workflow2

Molecular2

Assembly1

Nuclear transfer4

Streamline1

Sequence1

Study1

Greatly1

Innovations1

Dramatically1

Incomplete1

Development1

Research3

Human1