...both over- and under-expressing clones. If GFP-encoding cDNA is co-transfected at a favorable ratio to...Time CRISPR User) The Challenges of Cell Culture Plasmids 101: Mammalian Vectors Resources on Addgene.org...Our Protocols Pages Browse Fluorescent Protein Plasmids Find Mammalian Expression Vectors ...amount of agarose/DMEM should be proportionately increased). For some cells, add a drop of serum to aid in...up, most will eventually grow within a week. We screen the wells in groups as they become confluent, taking...important to have a functional assay in place to screen your clones. Depending on the protein, we have ...SH-SY5Y cells, as these cells did not seem to form discrete colonies but constantly migrated away from each...

... entire adult brain, using a plasmid that expresses a genetically encoded calcium sensor, GCaMP6 (Nguyen... eBook Read our plasmids 101 blog series Resources on Addgene.org Browse our plasmid collection pages...developing zebrafish pancreas. They first tagged these cells using plasmids. Then using HOTcre, a method that... resistant to chemotherapy. You can find these plasmids at Addgene. Another advantage of mice as model... feeding the worm bacteria expressing specific plasmids (L4440) that contain your gene of interest (Fire...al., 2015). If you are interested in C. elegans plasmids and resources check out Addgene’s Worm Expression...pages Read our molecular biology reference Find plasmids for your research ...

...Improvement of pCVD442, a suicide plasmid for gene allele exchange in bacteria." Plasmid 51.3 (2004): 246-255. PubMed...exogenous enzymes, require multiple transformation and plasmid curing steps, and can generate off-target mutations...a mutated allele introduced from a conjugative plasmid replaces the chromosomal copy. This is a versatile... These two steps are (Figure 1): The initial plasmid integration step (the so-called “single crossover...difficult, we have also included versions of pTOX that encode one of two visible blue or magenta chromoproteins... an allelic exchange experiment? Find the pTOX plasmids here! Continued development of new allelic exchange...Additional resources on the Addgene blog Read our Plasmids 101 blog posts Find our CRISPR blog posts Learn...

... phosphatase activity: In yeast, the PHO8 gene encodes a vacuolar alkaline phosphatase. Normally PHO8 ... Resources on Addgene.org Fluorescent Protein Plasmids and Resources Fluorescent Protein Guide: Biosensors...synthesized at the ER, delivered to the vacuole via the secretory pathway, and then cleaved to generate an active...In both cases, red fluorescence in the vacuole increased with longer exposure to autophagy-inducing conditions...

...that vectorization strategies include creating an AdV transfer plasmid with the E1 and E3 (Early 1 and Early...replication (Harrach, 2014). Given that the early genes encode proteins essential to initiating the whole process.... These genes’ functions are rescued by helper plasmids used during production and/or by genes present...made available by this deletion from the genomic plasmid can be used to house a recombinant sequence (transgene...late genes are usually present in the transfer plasmid for the majority of adenoviral vector systems used...

...Addgene blog:  Plasmids 101: What is a plasmid? Plasmids 101: Origin of ReplicationPlasmids 101: PCR...process known as plasmid multimerization. Plasmid Multimerization Early studies on plasmid topology recorded...ligate the plasmid, and re-transform the ligated plasmid to recover the monomeric form of the plasmid. Have...copy number plasmids causes instability: CoIE1 encodes a determinant essential for plasmid monomerization... monomer of Plasmid 12259: pMD2.G (depicted on the left) is ~5.8 kb in length. The plasmid can exist on... the sequencing results for their plasmids, they found the plasmids were much longer than expected, and...tandem. Such plasmids are known as multimers, and they are the result of multiple plasmid copies combining...

...Turquoise (CeNL; Plasmid #85199), Yellow (YeNL; Plasmid #85201), Orange (OeNL; plasmid #85202) and Red ... Researchers have been sharing plasmids ever since there were plasmids to share. Back when I was in the...specific strategies and tactics for getting the plasmids he wanted. Addgene makes it so much easier to ...kDa. An example is the CRISPaint-NLuc construct (Plasmid #67178) for use in the system outlined in Schmid-Burgk...CyOFP1-NLuc-CyOFP1; Antares). Direct injection of plasmids for Antares or firefly luciferase into mouse livers...Red (ReNL; plasmid #85203) NLuc fusions. The Nagai lab has additionally deposited more than 25 fusions of.... This construct is available through Addgene (Plasmid #83926). Live-Cell Voltage Sensor  Ingaki and ...

...page and can be used to create fusions with a gene of interest. These plasmids will be most useful for...transfection). Future of FRET The first genetically encoded FRET biosensor, Cameleon (Miyawaki et al., 1997...biosensors too, as another laboratory may have already created the sensor that you need. FRET biosensors designed...multiple FRET biosensors simultaneously would further increase the amount of correlated information available...

... Find CRISPR Plasmids for use in Mammals Check out Our CRISPR Guide Pages Find Plasmids for Mouse Targeting...model basics To create a mouse model, the gRNA, Cas9, and donor oligonucleotide or plasmid components are...genome is made up of 3 x 109 nucleotides (nt), and encodes 23,000 or so genes. It would be great if we could...target site, and a donor oligonucleotide or plasmid as the repair material if needed (for knock in models)...provided (in the form of an oligonucleotide or plasmid), precise changes can be made in the genome via...mice carrying the intended mutation. With a donor plasmid, the outcome is less predictable. In our best case...specifies the target, the Cas9 endonuclease that creates the DNA double-strand break (DSB) at the target...

...Non-coding RNAs (ncRNAs) are transcribed but do not encode proteins. They play important roles in many cellular...affect nearby genes (Figure 1A-C). Find CRISPR plasmids at Addgene! Knock-in: Introducing tags or stop...observe the effects. Luckily for us, CRISPR is an incredibly versatile system that can be tweaked to target...pairs long. Using a single guide RNA (sgRNA) to create a double-strand break works well because very small...ncRNA locus (Ho et al. 2012). For large-scale screening, sgRNA libraries are useful for targeting thousands...092902 Liu Y, Cao Z, Wang Y, et al. Genome-wide screening for functional long noncoding RNAs in human cells...