... efficiencies ranking as low, medium or high is based on the reprogramming efficiencies reported in González...Non-replicating Plasmids: Generating iPSCs with plasmid-based expression requires serial transfection of 1 or ...Replicating oriP/Epstein-Barr nuclear antigen-1 (EBNA1) based episomes: These plasmids carry the origin of replication...DNA methylation. Another downside of using EBNA1 based vectors is that the plasmid can hang around for ...behave. iPSCs are typically tested with molecular-based assays first. These methods less rigorously assesses...Transgene Expression Mediated by a Sendai Virus-Based Vector Using a Nonstructural C Protein and Endogenous...

...released report “Enhancing Dissemination of Evidence-Based Models for STEM PhD Career Development; a Stakeholder...development and implementation of collections of evidence-based practices. These concrete programs and practices...

... of DNA you need, and at which level of purity. Based on these premises we can classify a plasmid preparation...will adopt for the extraction. Restriction enzyme based approaches can require several hundreds of nanograms...

... your website? Many vendors will process orders based on a purchase order, which is an official document...depositing institutions. Addgene cannot fulfill orders based on PO documents that are sent to us by fax or email...

...resistance cassette) in pUC19 based entry vectors, as well as the pGreen-IIS based destination vectors. To ...CRISPR family of plasmids by offering a lentivirus-based mechanism for sgRNA delivery and providing a means...ACS Synth Biol 2013 Sep 20;2(9):537-41. DREADD-based chemogenetic technologies After several years...up to six DNA fragments at a time. This method (based on the Golden Gate technology) exploits the ability...reconstitution across synaptic partners) technique based on functional complementation between two non-fluorescent...tags in the FP sequence for separation from Cre-based Brainbow-expressing cells. Autobow plasmids are ...The expression plasmids for a given species are based on the same backbone to permit expression levels...

...resolution for in vivo work. APEX2, a faster peroxidase-based labeling approach, is an alternative option but ...signaling. Limitations of RaPID Like other BirA*-based proximity labeling methods, RaPID cannot distinguish...reported by other research groups that performed BirA*-based affinity purification mass spec experiments. These...

...protein degradation. Enter TRIM-Away - an antibody-based system developed by the James and Schuh labs. Clift...endocytosis is further complicated by the reliance on pH-based biosensors used to detect the maturation of neutral... Crabtree’s lab has developed a new CRISPR/Cas9-based system to study epigenetic modulation. While catalytically...

...recombination of the targeting vector. CRISPR/Cas9-based systems have been developed for generating knockin... for gestation. All three can introduce multiple base pair deletions, but CRISPR/Cas9 can also introduce...in which the conditional recombination can occur based on the cells in which Cre is expressed.  Several...

...is less than half the size of the Cas9 or Cas12a based systems, making it well suited for AAV packaging...was shown to be effective for CRISPR activation, base editing, and genome editing in mammalian cells. ...had the highest activity for gene activation  and base editing, whereas a different set of mutations (the...

...change, so they are typically used in microscopy-based assays like immunohistochemistry, though they are....  Biotin Biotin is used in a range of antibody-based assays. Biotin itself is not visible or capable ...But hopefully, this introduction provides a good base from which to dive deeper!References and resources...instruments, each other, and your target. I love FPbase’s Spectra Viewer tool for comparing different fluorophores...

...gradient with different concentrations of your protein-based buffer (FBS, milk, etc.) against the same amount...instead of concentrations. Most labs use protein-based buffers, so here’s a quick tip: Keep a bottle of...even give gelatin a shot.  Remember that protein-based blockers degrade fairly quickly, even at 4 °C, so...peroxidase protein (HRP), have a limited dynamic range based on the solution used to expose them. You’ll need...(if available) and note in analysis Plasmid-based proteins: plasmid sequence not accurate Check ...and/or concentration Try a different protein-based buffer (eg, milk instead of BSA) Nonspecific...buffer/adjust lysing buffer strength Sonication based lysing protocol Subcellular fractional kit or...

...diffusion rates. Smaller genetically encoded probes are based on cofactor binding proteins as described above....the His tag and several others. Another peptide-based strategy uses an enzyme to covalently label the ...: 9759496.  3. Buckley, Anthony M., et al. "LOV-based reporters for fluorescence imaging." Current opinion...PMC3129320.  18. Mastop, Marieke, et al. "A FRET-based biosensor for measuring Gα13 activation in single...protein are available (Cranfill et al., 2016 and FPbase.org). A number of variants remain fluorescent under...

...expression of many different genes was predictable based solely on the promoters and Shine-Delgarno sequences...Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end or paired-end...segments that can be engineered to bind specific RNA bases. By swapping out these different modular repeat ... the variant PUF segments required to bind any 8 base RNA sequence. The segments are provided in Gateway...

...Medical School/Brigham and Women’s Hospital (BWH). Based on her experience co-directing Mentor Circle Programs...-Europe professional collaborations, setting up bases and contacts with Spanish laboratories to start ...key to the success of this program. As we build a base of former mentees, we encourage them to get involved...consulting firm focused on new health care models based on integrative and eco-social health strategy, and...their degree. Students filled in a pre-program, baseline self-assessment survey to determine their expectations...expectations of the program and their baseline skills. Mentors were Spanish scientists (MD, PhD and MD/...defining expectations of the program and their baseline skills and a post-program self-assessment, participants...

...difficult, but other tissues like the retina have cell-based therapies underway. Chapter 7: Generating entire...entire lungs using a decellurized scaffold as a base. Darrell hopes that further advancement of this technique...

... after I arrived, I was looking through a paper-based, affordable, and portable microscope called the ... known as gas vesicles) as acoustic biomolecule-based dyes. This technology is widely applicable as bacterial...

... predict. Slower Dephosphorylation Rates: FRET-based reporters can have slower dephosphorylation rates...occurs in the cytoplasm and/or nucleus, while FRET-based reporters can be used to measure kinase activity...

... you. Traditional restriction enzyme and ligase based cloning is a relatively inexpensive way to get the...a try. This method, developed by Invitrogen and based on lambda phage integration sequences, relies on...inserts a breeze. Among them are some of the homology based methods, like Gibson or SLIC, since one can pool...

... that need them. Adeno-associated virus (AAV) – based delivery techniques may hold the key. In the simplest...effective. Challenges for AAV gene therapy Although AAV-based gene therapies are promising, scientists are working...should be taken into account when developing AAV-based therapies. Furthermore, deeper understanding of ...

...Francois St-Pierre’s lab developed a new form of FACS-based cell screening, Single-cell Phenotypic Observation...Plasmids and Resources page.  CDDR (CRISPR–Cas9-based Dual-fluorescent DSB Repair) is a double-strand ...double-strand break repair assay for mammalian cells. It's based on the introduction & resolution of two DSBs in a...eliminate competing mobile genetic elements. Image by Basem Al-Shayeb in the Doudna Lab. Scientists looking...