... and ligated into your desired plasmid. It is important to note that restriction enzyme target sites can...be found throughout DNA sequences. Thus it is important to confirm that there are no additional sites ...complementary to the free nucleotide. Given this opportunity, T4 will resume its polymerase activity, add ...repaired by the normal replication process. It is important to note that LIC has difficulty assembling DNA...

...can help you begin to understand what genes are important to a certain phenotype, and allow you to design...the screening process. Amplify the library (electroporation and maxiprep). If delivering as virus, make...die or do not pass selection (in the case of a reporter) Sort “winning” cells Sequence those cells that...

...γ-retroviruses differ from lentiviruses? There are a few important differences between engineered γ-retroviruses ...small enough to enter the nucleus through nuclear pore complex. This feature may be useful for the transduction... expression of transgenes via increased nuclear export. LTR Subcomponents: U3 R U5 in cis LTR; Long terminal...

...silencers. Promoter Regions There are three main portions that make up a promoter: core promoter, proximal...promote the initiation of transcription. The most 3' portion (closest to the gene's start codon) of the core...transcription factors bind. Distal Promoter The final portion of the promoter region is called the distal promoter...

...lentiviruses integrate into the genome. This is important because local chromatin structure at the site ...

...customized fluorescent reporters can be challenging enough, but incorporating multiple reporters into a single ... the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP) the reporter mRNA, which contains...their reporter mRNAs. Using the TRICK plasmids, Halstead et al demonstrated that 91% of reporter mRNAs... used to study transport characteristics of various Golgi and plasma membrane reporters as well as secretory... protein transport. Franck Perez’s lab has developed a whole collection of hook and reporter proteins ...pyralis) reporter plasmids for use in high-throughput screening of pre-mRNA splicing. This reporter system...widely used luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids have been...

...the pSer-incorporated polypeptide. These three components were shown to effectively incorporate phosphoserine...Creating phosphoserine-incorporated proteins The generation of pSer-incorporated proteins requires a molecular...strain for pSer-incorporation The second component necessary for phosphoserine incorporation is a modified...considerations, it is important to determine how much phosphorserine has actually been incorporated into the full-length...different levels of incorporation due to differences in stability of the p-Ser incorporated protein as well...determine the stoichiometry of phospho-incorporated to non-phospho-incorporated proteins purified from the system...pSer-incorporated to non-phosphorylated protein. Verification of site-specific pSer incorporation can ...

... facilitate transport: retrograde transport relies on dynein, while anterograde transport relies on kinesin...take advantage of existing transport pathways in neurons. Anterograde transport is typically used for the... transmitter synthesis. Retrograde transport is used for transport of endocytosed material or molecules... impaired, and only few reports exist about dye molecules that are transported from one neuron to another...There are several reports of both retrograde and anterograde intracellular transport of some AAV serotypes...including dendritic and axonal extensions and, most importantly, any connections the neurons make to other distant...for various forms of retrograde and anterograde transport suggest that there are several parallel mechanisms...

... whether respondents saw them as not important to very important. In addition, we also explored the barriers...”. The study also showed that It is important that an ELN supports all types of data, which enables researchers...electronic ones? It is important to know in which ways your data can be exported or printed from the ELN...project. Can you meet the ELN support team? We cannot stress enough how important this is when deploying an...only 46% could be reproduced. In a recent Nature report from researchers at Amgen, it was stated that only...through all saved data and automated writing of reports. To be able to understand the situation in depth... is likely to require on-going maintenance and support.” Besides that, academic institutions tend to think...

...conjugated with a reporter or tag that can then be detected.  Typically, the reporter is an enzyme such...indirect and sandwich ELISA the reporter signal is directly proportional to the level of antigen in the...whereas in a competitive ELISA the reporter signal is inversely proportional to the level of antigen in the...dish and incubate it with a reporter conjugated primary antibody. The reporter reacts with a provided substrate...simplicity we will discuss the more common enzyme-based reporter methods. For enzymatic signal detection, users... sandwich and competitive. In a direct ELISA a reporter enzyme conjugated antibody binds to antigen coated...antibody first binds to the antigen and then a reporter enzyme conjugated secondary antibody binds to ...

...you think there are four important times to measure that growth rate, you report the results of all four... registered reports, the gold standard for testing hypotheses Finally, Registered Reports combine all ...Reproducibility Project: Cancer Biology, where reported methods may not include how many tests were conducted...attention on the results instead of the methods. Unreported flexibility and questionable research practices...result after conducting a replication study. An important difference between the original setting and the...genetic variation, or different handling can all be important.  If we eliminate the problems above: unclear ...research finding we can be more certain that some important, previously unknown variable actually makes a ...

... to cleave and activate a reporter. The detectable signal from this reporter can be measured and quantified...and a Csm6 specific reporter (in the same fluorescent channel as the Cas13 reporter), they could significantly...variants, Cas13, and Cas12a, to develop simple, portable, and inexpensive platforms to reliably detect ...method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK and SHERLOCKv2) (Gootenberg... termed DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) (Chen et al., 2018). Both SHERLOCK and... SHERLOCK- Specific High Sensitivity Enzymatic Reporter UnLOCKING Cas13 (C2c2) was first identified in...molecules. In SHERLOCK a quenched fluorescent ssRNA reporter is added to the reaction. Cleavage of the quenchable...

... the 3’ UTR of the reporter mRNA via an MS2 coat protein (NLS-MS2-RFP) the reporter mRNA, which contains...removal of GFP from the TRICK reporter. Delayed translation is known to be important in development, and Halstead...proteins to allow translation to occur. For a TRICK reporter mRNA this displacement leads to a change in the...GFP protein that binds to the coding region of a reporter mRNA via a PP7 coat protein (NLS-PCP-GFP) an RFP...translation, both GFP and RFP are bound to the reporter, causing it to appear yellow. The act of translation...of the mRNA, leaving only a red FP bound to the reporter. Because it contains a nuclear localization signal...copies of the FP-coat protein fusions to their reporter mRNAs. Using translation inhibitors cycloheximide...

...the promoter of the reporter gene to activate expression (Figure 2c). The reporter gene then allows detection...genes as reporters and some that repress instead of activate transcription. The HIS3 reporter system (...sequence (UAS) of its target gene, in this case a reporter gene (e.g. luciferase or GFP), regardless of whether...co-expressed in a yeast strain that also contains a reporter gene cloned downstream of a promoter containing...referred to as the “bait”, Figure 2b) cannot activate reporter gene expression on its own, but, if the two POI...Gal4 activator, bacterial LexA DBD and the lacZ reporter gene (3, 4), there are now many variations, including...to have Y2H systems that avoid transcriptional reporters altogether; In the split-luciferase system, intein-mediated...

...cuts to important research funding.  So when I saw that there was going to be a march in support of science...the rain, about 4,000 supporters of science showed up to march. We had full support of local law enforcement...proved to be the perfect opportunity to cement in my children's heads the importance of science and positively...seems to break everyday. Many researchers and supporters of science marched to advocate for scientist ...for science, and numerous other reasons. It is important to remember that we, scientists and non-scientists...°W – Vanessa Sung Hundreds of scientists and supporters in Montreal took to the streets on April 22 to...Albany, Rep. Paul Tonko also came and spoke in support of science - he's one of the few members of Congress...

...has dubbed these fusions LumiFluor Reporters. The LumiFluor reporter constructs were designed to be packaged... The researchers took a FRET reporter and adapted it into a BRET reporter by using NanoLuc® Luciferase...share your constructs from lab to lab. Promega supports the Addgene mission statement: Accelerate research...(FLuc) and Renilla (RLuc) luciferase. This is important not so much for how bright you can make a reaction...couples NLuc to fluorescent proteins to make better reporters and the second couples NLuc to fluorescent proteins...host cells. The fluorescent moiety of the BRET reporter was used to sort successfully infected/expressing...NanoLuc luciferase. Both GpNLuc and OgNLuc BRET reporters produced bright signals without exogenous excitation...

... to report mitochondrial turnover Mitochondrial health, transport and turnover are important features...genes, reporters, or other markers into a target genome. They did this by selecting the important elements... be imported to the mitochondrial matrix. Gottlieb’s lab has used an inducible MitoTimer reporter using...usefulness in cell culture to report changes in mitochondrial turnover and transport. pMitoTimer from Yan’s ...s lab, which is constitutively expressed, reports on the balance of biogenesis and mitochondrial degradation...mitochondria in the networks marks the differences in import of newly synthesized (green) protein and organelles...channelrhodopsins. CnChR1, nicknamed Chrimson, is the first reported yellow-peaked channelrhodopsin with a spectral...

...mentor Your manager should support you, just like you are supporting your report(s). This means that you ...unique skills and strengths (you will likely have reports with different levels of experience), find out ...how to prioritize but I  just added the “most important” tasks my schedule. This was bad for 2 reasons...tasks before I was a manager! Second, my direct report was not engaged because I only left him/her with...expectations, answer any questions, and (most importantly) decide on a timeline. My experience is that ...you’re delegating a recurring task to a seasoned report, there should be questions! Remember that delegating...develop your team members by giving them new opportunities. I still need to check myself sometimes to ...

...of the opportunities you’re provided throughout your PhD program.   Find people who will support your ...had a wonderfully supportive mentor and graduate office. I cannot stress how important they were to my ...experimenting with the career and networking opportunities available to me as a graduate student culminated...events for adults and professional development opportunities for researchers interested in connecting with...they had my back. If your advisors are not as supportive of your career goals, find people who are. I ...as types of experiments. I searched for new opportunities with an open mind and open ears. Everyone I ...mentioned, my graduate office staff was amazingly supportive. I talked to them about my interests and my ideas...

...everywhere can access plasmids despite the evolving export/import rules. Our shipping and lab teams continue to...entire supportive Board, we have a robust committee process in place and continue to incorporate additional...all of the success we have achieved and the opportunities we have for the future. At Addgene we will never... The success of these projects allows us to support more new projects. Diversification is happening... quality and success. Customer and technical support Our customer and technical service brands continue...sharing, but we don’t have to worry about getting support from depositors. We regularly hear how much they...blessed with a large community of scientists who support us by depositing, requesting and spreading the ...