...Gene Expression Analysis During iPS Reprogramming Process. Sci Rep 8, 8716 (2018). https://doi.org/10.1038...variable nonsense to publication quality data. Sci Rep 7, 2409 (2017). https://doi.org/10.1038/s41598-017...ensure you’re recording and reporting the right information so others can reproduce your efforts. And if you...step. Scorpion chemistry in particular has been reported to work well in the Lightcycler while being relatively...Yes Agarose gel Sunrise primers   Low Reporter and quencher on hairpin structure included in ...much as sixfold variation in expression levels reported (Biassoni, 2014), so you’ll want to ensure that...steady pair of hands to produce reliable and reproducible data. Here are some tips to help you in your...

...bfloGFPc1. Bomati et al., Sci Rep 2014 Jun 27;4:5469. MitoTimer plasmids to report mitochondrial turnover ...the cells will turn green. Mashiko et al., Sci Rep 2013 Nov 27; 3:3355. Rinehart lab reagents for...protocol for TALEN construction. Sakuma et al., Sci Rep. 2013. Nov 29;3:3379. Interested in more genome...and 13 in the TALE repeat, other naturally-occurring variations in the TALE repeat, referred to as “non-RVD...an identical variable repeat (VR) at the 4th and 32nd residues of the TALE repeat and the VR differs between...mutagenesis libraries or to insert heterologous genes, reporters, or other markers into a target genome. They did...Gottlieb’s lab has used an inducible MitoTimer reporter using a Tet-on system (pTRE-Tight-MitoTimer) to...

...for multiplexed super-resolution microscopy. Cell Rep Methods 2(10):100301. https://doi.org/10.1016/j.crmeth...by reducing the distance between the target and reporter. They have fewer non-specific binding opportunities...it either needs to be directly conjugated to a reporter or you need a light-chain specific conjugated ...fairly easy to find these in a Fab fragment format. Reporter-conjugated anti-IgG Fab fragments make good secondaries...

...Monoclonal Antibody Sequences. Sci Rep 6: . https://doi.org/10.1038/srep31730 Tsuruta LR, dos ML, Moro AM (...recombinant antibodies are essential for good reproducibility. Non-recombinant antibodies are notorious for... for being the source of irreproducible data (Begley & Ellis, 2012; Baker, 2015). This contributes to ...doi.org/10.7554/elife.43322 Baker M (2015) Reproducibility crisis: Blame it on the antibodies. Nature ...10.1038/483531a Bradbury A, Plückthun A (2015) Reproducibility: Standardize antibodies used in research. Nature...

...question may take some time. If the customer service rep does not tell you upfront how long it will take to...

...for high-titer lentivirus preparations without ultracentrifugation. Sci Rep. 2015; 5:13875. PubMed PMID...prevent the formation of replication competent lentivirus (RCL). Replication competence can arise from... addition, the promoter of the 5’ long terminal repeat (LTR) is mutated in third generation systems such...considerably lower volume than with the unconcentrated prep. For example, at Addgene we typically concentrate...concentrate 100-fold meaning that a 10 cm2 dish preparation of virus will only yield 150 μl of material as opposed...opposed to the 15 ml collected from an unconcentrated prep. With this in mind, if the experiment requires ...

...diverse array of human cells. Sci Rep 4: . https://doi.org/10.1038/srep05105 Stephens CJ, Lauron EJ, Kashentseva...Scientific Reports paper introducing the delivery method in 2014, Manuel Goncalves lab report that AdV-...provide a DNA template to repair the double strand break via homology directed repair and you would probably...years for in vivo delivery and gene therapy and represent one of the most studied viruses. Adenoviruses ...utilized the Non Homologous End Joining (NHEJ) DNA repair mechanism to knock out genes as a result of an ...approach to NHEJ is to utilize the Homology Directed Repair (HDR) to knock in corrective genes. For this you...for any such disease. This technology therefore represents a potential platform that could be realized via...

...Degron Tagging with Short Homology Donors.” Cell Rep.15(1) (2016):210-8. PubMed PMID: 27052166. Kunzelmann...including protein enhancers and repressors, can be included in the repair template for inducible alteration...overexpressed fluorescent protein fusions is to replace the genomic copy of your gene of interest with ...site and, using the endogenous homology directed repair pathway, a new user-defined DNA sequence (like ...to individually generate gRNAs and homology arm repair templates containing the tag for each insertion...sequence, as well as the 5’ and 3’ homology arm repair template (Fig. 1, Step 1). There is no need for...expression and the second site accepts the homology arm repair template. SapTrap includes a prebuilt donor plasmid...

...bp, plus a 20 bp barcode 400 ng Solid pCMV-Rep 78/68 SSM Library Amplification and Sequencing Protocol...amplification protocol with your deposit. This is because repeated amplifications of pooled libraries can lead to...containing smaller, recombined backbones that may replicate faster. For this reason, if your library has inserts...recommend ensuring the cells used are meant for repetitive sequences (such as STBL4 or Endura Duo), growing...Protoc. 2017 with modifications Screening pCMV-Rep78/68 Scanning Saturation Mutagenesis (SSM) Pooled ...Addgene blog Genome-Wide Screening Using CRISPR Preparing Pooled Libraries NGS Quality Control for Pooled...

...fluorescent voltage sensor to detect neural activity. Sci Rep 9, 15878 (2019). doi: 10.1038/s41598-019-52370-8 ...sensors are better classified by their fluorescent reporter and the mechanism of their voltage sensing component...

... the SAM system differ in target preference. Sci Rep 9: . https://doi.org/10.1038/s41598-019-54179-x Tanenbaum...single copy of VP64 per each dCas9, SunTag uses a repeating peptide array to fused with multiple copies of...A, Zhang F, Marraffini LA (2013) Programmable repression and activation of bacterial gene expression using...Doudna JA, Weissman JS, Arkin AP, Lim WA (2013) Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific...

...Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: 26456817. Find plasmids from...methods for repairing DNA after a double-stranded break. HDR copies the sequence from a repair template ...HDR, and MMEJ repair varies from organism to organism. MMEJ doesn’t yield a “perfect repair” like HDR, but...end-joining (NHEJ) to make deletions or homology-directed repair (HDR) to create precise genome edits. But have ...have you heard of another double-stranded break repair mechanism: MMEJ (microhomology-mediated end-joining... only very small homology regions (5-25 bp) for repair, making it easier to construct targeting vectors...with flanking sequence homology for error-free DSB repair. NHEJ joins the ends of a DSB in an error-prone...

...·U wobble base pair. EMBO Rep 1:18–23 . https://doi.org/10.1093/embo-reports/kvd001 Additional resources...even been linked to some diseases. It has been reported that 5-10% of human genes contain a region where...

...performed using the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding...through how we produce both our AAV preps and our lentiviral preps.   Viruses are generated using standard...order to generate high quality preparations. After production, all virus preps are titered and subjected to...-containing) AAV particles within representative vector preparations was quantified with electron microscopy... lentiviral preparations (“-LVC” catalog items) are generated from lentiviral preparations (described ... to the cleared viral preparation pool. In addition, the final viral preparation undergoes PCR with primers...Addgene has a growing catalogue of viral preps available to request. In this post, we’ll walk you through...

...Gomez-Nicola D, Riecken K, Fehse B, Perry VH. Sci Rep. 2014 Dec 22;4:7520. PubMed PMID: 25531807. PubMed...desired transgene flanked by HIV-1 long terminal repeats (LTRs) that facilitate viral packaging and host...transduced rat neurons in vivo with beta-gal and other reporters, and they showed that expression persisted for...original system, the possibility of creating a replication-competent virus via recombination between the...of recombination events necessary to create a replication-competent virus. Second, Tat is eliminated through...

... 2. Replicates and repeats—what is the difference and is it significant? Vaux DL, et al. EMBO Rep. Apr...same aliquot/preparation. The replicate control isn’t measuring or assessing reproducibility of the effect...independent variable. Replicate controls: technical and biological Reliable results are reproducible from properly...two types of replicate controls: technical and biological. In general, technical replicates can be thought...Conversely, biological replicates ARE independent and can be thought of as "reproducibility controls". Biological...several different days. Biological replicates control for the reproducibility and consistency of the observed... employ transfection, negative, positive, and replicate controls.  Transfection controls: empty vector...

...Saito K, Ohki R, Fujii H. Sci Rep. 2013 Nov 8;3:3171. doi: 10.1038/srep03171. Efficient isolation of specific..., as well as the Editorial Board of Scientific Reports and PLoS One. Additionally, you are involved in...

...compact expression cassettes. Sci Rep 5: . https://doi.org/10.1038/srep10777  Garneau JE, Dupuis M-È, Villion...CRISPR-based RNA editing, the REPAIR (RNA Editing for Programmable A to I Replacement), system is also small ...infection of certain tissues. However, packaging Streptococcus pyogenes (SpCas9) and a gRNA together (~4.2 ...Cas9 include the use of St1Cas9 (~3.3 kb) from Streptococcus thermophilus and a rationally-designed truncated...direct in situ breaks labeling, enrichment on streptavidin and next-generation sequencing). Using this ...

...allow for replication. However, if plasmids have their own replication machinery such as Rep or other ...ori2, ori3) and replication initiation proteins (RepA, RepB) necessary for replication in various hosts... and W are known to have replicons with broad host ranges. Popular replicons from these groups include...different hosts that a plasmid can transfer to, replicate in, and persist in. Some plasmids have a narrow... while others have a broad host range and can replicate and stably maintain the genes they carry among.... These features are: The plasmid origin of replication (ori) contains structural elements that are versatile...structure allows plasmids to utilize diverse host replication machinery. For instance, the ori typically contains...

...longer single-stranded DNA.” Sci Rep. 5 (2015):12799. DOI: 10.1038/srep12799 PubMed PMID: 26242611 PubMed ...ll design your repair template with 55-100 bp homology on each side. For optimal repair, place the loxP... of ssDNA donor molecules for homology directed repair. Using long ssDNA donors, the Gurumurthy and Ohtsuka...suffer from low efficiency when a standard dsDNA repair template is used. The original design for creating...two gRNAs targeting two intronic regions and two repair templates in order to insert two loxP sites. Since...including one loxP site insertion and one NHEJ-repaired indel, often occur. Figure 1: In conventional...courtesy of Wenning Qin. Single-stranded DNA repair templates Short single-stranded DNA templates have...