We narrowed to 511 results for: plasmid dna
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TypeBlog Post... How to Use the Nanobody Toolbox Find Nanobody Plasmids Structural comparison As the name suggests, ... Species Antibody Subclass Nanobody Addgene Plasmid Rabbit N/A* TP897 pTP1183 (1x cysteine) pTP955...Nanobody In Bacteria: Table 2 also has a link to the plasmid you’ll need to express your nanobody of choice ...column. bdNEDP1 protease can be generated using plasmid pDG02583. Label the Nanobodies: Purified nanobodies...Journal of Cell Biology. PubMed PMID: 29263082 Find plasmids from this publication at Addgene. 2. Bates, M...23495938. Additional Resources on the Addgene Blog DIY DNA ladders from Penn State University Use Nanobodies... on Addgene.org Find more nanobody expression plasmids here Additional Resources Looking for monoclonal...
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The AAVantages of AAV in CRISPR Screens
TypeBlog Post...episomal expression. Episomes are circular pieces of DNA that reside outside a host cell’s genome. In this... The most obvious workaround is to use a multi-plasmid system, where you have one AAV vector dedicated...levels of AAV vector integration into CRISPR-induced DNA breaks. Nature Communications, 10(1). https://doi.org... -
AAVs in Retinal Gene Therapy
TypeBlog Post...random) integration into the genome, and its large DNA packaging capacity is well suited for larger transgenes...AAV. Its main limitation is the small size of its DNA cargo - 4.8 kb at most - which means that a gene ...in mammals Resources at Addgene.org Find AAV plasmids Find ready-to-use AAV viral preps Browse all viral... -
New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries
TypeBlog Post...derivatives. Unmodified Cas9 causes double stranded DNA breaks that commonly result in a complete loss of...These mutations result in an inactive RNA-guided DNA binding protein (dCas9). dCas9 is then tethered to... CRISPR Resources on Addgene.org Find CRISPR plasmids and resources here Find pooled libraries available...viability genes as they are subject to an increase in dsDNA breaks that can cause cell death. Thus, CRISPR inhibition...mitigating toxicity induced by high copy number dsDNA cutting. A Dolcetto library containing only 3 sgRNAs... -
The Challenges of Cell Culture
TypeBlog Post...good that it is used as a forensic technique (aka DNA fingerprinting) that can place suspects at the scene...range of testing, from animal doping to forensics, DNA sequencing to drug testing. You can find Nick on ...Protein Localization Resources on Addgene.org Find Plasmids for Mammalian Cells ... -
FlipGFP, a novel fluorescence protease reporter to study apoptosis
TypeBlog Post... external signals such as developmental cues and DNA damage. These signals activate a cascade of caspases...these diseases and disorders. You can find the plasmids for FlipGFP and FlipCherry at Addgene. References...Download the fluorescent proteins 101 eBook Read our plasmids 101 blog series Resources on Addgene.org Browse...protein fusions Browse the fluorescent proteins plasmid kits ... -
Plasmids 101: The Promoter Region – Let's Go!
TypeBlog Post...Once you've gotten to know what a plasmid is in general, it's time to take a look at some of its parts...origin of replication are necessary to replicate plasmids and make sure the cells maintains it. Here, we'll...the promoter, which is necessary for getting the plasmid express our gene of interest. This element is responsible... The RNA polymerase(s) RNA is transcribed from DNA using an RNA polymerase (RNAP). In bacteria this .... This means that the promoter present in your plasmid backbone must to be compatible with the type of...transcript, you will also need to make sure your plasmid has a promoter suited to working in your host organism...development-specific promoters available to scientists. Plasmids are oftentimes put to therapeutic uses, and in... -
Viral Vectors 101: Producing Your rAAV
TypeBlog Post...packaging plasmids: the pHelper plasmid containing Ad E2A, E4, and VA RNA genes; and the pRepCap plasmid containing...to be digested with DNase before undergoing titering, to remove residual plasmid DNA carried over from ...with your transfer plasmid, carrying your gene of interest, and the two helper plasmids, commonly called...called “triple plasmid transfection” (Figure 1). Figure 1: Overview of key steps of AAV production, iodixanol...repeats (ITR). You should always have a high-quality plasmid standard and rAAV reference material of a known...blog Parts of the AAV Packaging PlasmidParts of the AAV Transfer PlasmidAAV Variables That Matter AAV protocols... -
Cre-Lox and Other Site-Specific Recombinases
TypeCollection...Site-Specific Recombinases Cre Plasmids Flp Plasmids Dre Plasmids Other Recombinase Plasmids Recombinase-Dependent... cleave all four DNA strands at once. They can mediate the integration of a large DNA payload containing...same DNA strand in opposite directions, recombination results in an inversion and the region of DNA between... circular piece of DNA (and is not maintained). C) If the sites are on separate DNA molecules (in trans...recombination of the DNA between target sites. Codon optimization: Modifying the DNA sequence of the recombinase... vectors. Highlighted Plasmids Browse the tables below to find popular plasmids expressing your favorite...tamoxifen”, or “light”): Cre Recombinase Plasmids ID Plasmid Description Vector Type Promoter PI Additional... -
CRISPR Plasmids - Yeast
TypeCollection...cytosine base editor that can alter DNA bases without inducing a DNA break. Cytosine base editors convert...gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter...gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene/Insert Promoter... purify a region of genomic DNA and its associated proteins, RNA, and DNA. The enCHIP system uses an anti-FLAG...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Validated In...cerevisiae LEU2 none, need Cas9 plasmid Lu Do you have suggestions for other plasmids that should be added to ...changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and a... -
CRISPR Plasmids - Cascade-Cas3
TypeCollection...family. After nicking the target DNA, Cas3 initiates single-strand DNA degradation. This degradation can...CRISPR-Cas3 plasmids take an all-in-one approach and include Cas3 and Cascade on one single plasmid, though... the tables below for CRISPR Cascade-Cas3 plasmids. Plasmids are available for expression in mammalian...plants. Mammalian ID Plasmid Gene/Insert Promoter PI Publication Bacteria ID Plasmid Gene/Insert Promoter...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...Find plasmids for CRISPR-Cas and Cascade for large genomic deletions. CRISPR... CRISPR Cas3 CRISPR Plasmids: Cascade-Cas3... -
Protocol - pLKO.1 – TRC Cloning Vector
TypeProtocol...measure the DNA concentration, use 1 μL) 2 μL 10x NEB T4 DNA ligase buffer 1 μL NEB T4 DNA ligase add ...transfect plasmid DNA directly into the target cells. The shRNA will be expressed, but the DNA is unlikely...μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging plasmid 250 ng pMD2.G envelope plasmid to 20 μl serum-...to vary the ratio of shRNA plasmid, packaging plasmid, and envelope plasmid to obtain the ratio that gives...shRNA. psPAX2 Packaging plasmid for producing viral particles. pMD2.G Envelope plasmid for producing viral... also be used with packaging plasmid pCMV-dR8.2 dvpr and envelope plasmid pCMV-VSVG from Robert Weinberg...#B7001S NEB buffer 2 NEB #B7002S T4 DNA ligase NEB #M0202S T4 DNA ligase buffer NEB #B0202S DH5 alpha ... -
Protocol - How to Inoculate a Bacterial Culture
TypeProtocol...Storage of Plasmids Recovering Plasmid DNA from Bacterial Culture Background Information Plasmids can carry...isolate your plasmid DNA from the bacterial culture by following Isolating Your Plasmid DNA . Antibiotic...copy regardless of plasmid size. See the plasmid's info page to determine if your plasmid is high or low ...resistance gene on a plasmids allows researchers to easily isolate bacteria containing that plasmid from bacteria...numbers of bacteria necessary to isolate enough plasmid DNA for experimental use. The following protocol ... low copy plasmids? The copy number refers to the number of copies of an individual plasmid within a single...number plasmids should only need to be grown for 12-16 hr on average. Certain features of a plasmid may ... -
CRISPR History and Development for Genome Engineering
TypeCollection...posts , or find CRISPR plasmids for your research. Addgene has CRISPR plasmids for the following models...editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature . 533(7603):420-4....Fonfara I, Hauer M, Doudna JA, Charpentier E. 2012. A programmable dual-RNA-guided DNA endonuclease in adaptive...specifically, specialized Cas proteins snip foreign DNA into small fragments approximately 20 bp in length...length and paste them into contiguous stretches of DNA known as CRISPR arrays. Separate Cas proteins then...motif (PAM). The CRISPR complex binds to the foreign DNA and cleaves it to destroy the invader ( Figure 1 ...protospacer sequences that have homology to foreign DNA. Protospacers are separated by short palindromic ... -
Adapting Toehold Switches to Detect Zika Virus
TypeBlog Post... Check out All the Plasmids from the Collin's Lab Find Synthetic Biology Plasmids for Your Next Experiment... days of lost productivity waiting for follow-up DNA oligos to arrive. We also were careful to set up ...Components." Cell (2016). PubMed PMID: 27160350. Find Plasmids from the publication at Addgene 2. Green, Alexander...Experiment Deposit Your Plasmids with Addgene ... -
Plasmid Modification by Annealed Oligo Cloning (with Protocols)
TypeProtocol...Restriction Digest of Plasmid DNA DNA Ligation Bacterial Transformation Summary Oligo overlap cloning can...anytime you need to add a short stretch of DNA to a plasmid, such as: Changing a Multiple Cloning Site...cut out the band containing your vector DNA. Gel purify your DNA away from the agarose using a commercially...Modify your plasmid MCS or add short sequences using oligo overlap cloning. Protocols... Protocols Plasmid Modification...Modification by Annealed Oligo Cloning Plasmid Modification by Annealed Oligo Cloning You may also like... Restriction... -
A Deep Dive into BioBuilder
TypeBlog Post...Standards Find Plasmids for Creating and Controlling Gene Networks Browse All SynBio Plasmids ...cellular extracts as a means to rapidly prototype DNA circuits and logic gates. This BioBuilder-Caltech... -
TALEN Guide
TypeCollection... “multiple DNA fragments in an ordered fashion in a single reaction.” Using these plasmids, which are ...Guide to TALEN technology and using TALEN plasmids TALENs...Technologies You may also like... TALEN Kits CRISPR Plasmids Zinc Finger Consortium Originally published in...Imagine being able to manipulate a specific region of DNA in the genome of your favorite species almost as ...biological disciplines are coming closer to this DNA targeting ideal. Figure 1: (A) Representation of ...enzymes and other useful protein domains to a specific DNA sequence ( Figure 1A ). Scientists have identified...expect that assembling relatively small, repeat-laden DNA regions into a single construct would be technically... -
New Tools Enable CRISPRa for Neuroscience Applications
TypeBlog Post...comprehensive CRISPR guide Find CRISPR plasmids and resources Find CRISPRa plasmids ...genes can be relatively laborious because multiple plasmids/viruses are needed to deliver all the protein ...construct for generating the SPH mouse. (B) AAV plasmids used to express CRE under neuron-specific promoters...individual neuronal populations. The Yang Lab’s AAV plasmids used to express Cre under neuron-specific promoters... CD8+ cells were then isolated and their genomic DNA sequenced. Guide RNAs targeting several known and...sensitive applications such as CRISPR screening. The plasmids used by the Qi Lab for this particular application...simpler to implement in genome-wide screens over cDNA libraries. However, using CRISPRa to target individual... -
Cpf1: A New Tool for CRISPR Genome Editing
TypeBlog Post...Cas9’s second HNH endonuclease domain. Cpf1 cleaves DNA in a staggered pattern and requires only one RNA ...Francisella Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover FnCpf1’s PAM sequence...system." Cell (2015). PubMed PMID: 26422227. Find plasmids from this publication at Addgene. 2. Zetsche,....1 (2016): 31-34. PubMed PMID: 27918548. Find plasmids from this publication at Addgene. 3. Kim, Daesik...Biotechnol. (2016). PubMed PMID: 27272384 Find plasmids from this publication at Addgene. Kleinstiver...Biotechnol. (2016). PubMed PMID: 27347757. Find plasmids from this publication at Addgene. Makarova, Kira...PMID: 26411297. Resources at Addgene Find CRISPR plasmids at Addgene Read Addgene's CRISPR Resource Guide...