...be translated to other lines. Differences in the physiology of different cell types, however, can make...not only between serum from different manufacturers but also between different lots of serum from the same...low levels of contamination. For those labs that prefer a more economical mode of detection, Uphoff et....the second. When harvesting lentivirus, some labs prefer to collect and pool the harvests from multiple ...for a single harvest after transfecting with the transfer and packaging plasmids. A benefit of the multiple... as the transgene expressed, the viral storage buffers used, and other experimental parameters. For example...concentrated dilutions you will be able to overcome differences in receptor expression. It's all in the mix Even...

...dopamine sensors offer different levels of apparent dopamine affinity, with 113050 offering high apparent...these readily available to the community. We now offer AAV9 for 2 more constructs from Christopher Harvey... various AAV serotypes can be used to achieve different patterns of expression, researchers often need...empirically for their own experiments. To this end, we offer specific control AAV in smaller aliquots of 20 ul.... The Addgene viral service has now expanded to offer many of our control plasmids in new serotypes. See...sensitive response to action potentials, is now offered under control of the CAG promoter for ubiquitous...monitoring of neurotransmitters and we are excited to offer AAV for the newest GRAB sensors from the Li Lab....

...converted to a different isotype, then two different secondary antibodies can be used with different fluorescent...have different binding properties (Morelock 1994). While in the majority of cases the differences are ...conversions and chimeric antibodies! References and resources References Liddell, E. (2013). Antibodies. In...changing an antibody’s natural isotype to that of a different group (Fig. 1). This process, called isotope conversion...antibody’s isotypes, however, you could then use 2 different secondaries for your antibody pair and visualize...constant regions or solely the Fc region of a different species and/or isotype (Liddell 2013). In most...

...experiment uses 2 or 3 different fluorescent colors each measuring a different target. Flow cytometers...analysis. Since different populations of immune cells express unique Clusters of Differentiation cell surface...flow cytometry experiments.       References and resources References: Bonner WA, Hulett HR, Sweet RG, ...  Cells are first resuspended in a pressurized buffer called sheath fluid and transported through tubes...each cell as it passes through and scatters in different directions. The degree of scatter is captured ...highly versatile and some can accommodate up to 30 different colors. This lends flow cytometry particularly...to complex experiments looking at a variety of different targets. If you are planning on using fluorescent...

...isotype is expressed at different times of the immune response and initiates different immune cascades. In...generates an antibody response in the immune system is referred to as an antigen. You can learn more about antibodies...In the research setting, antibodies of different isotypes can be used together in the same experiment ...of epitopes on the target protein, so they are referred to as polyclonal. Polyclonal antibodies have the...However, each animal will make antibodies against different protein epitopes and so variability between lots...molecular weight by SDS-PAGE, and are subsequently transferred to a membrane. A primary antibody recognizing...dissociated from the bead and the antibody using a low-pH buffer. Then, the purified protein can be used in experiments...

...directions in your research. We all have our preferences. Your preferred type of CRISPR will likely depend on ...horizon (Altae-Tran et al., 2023)! References and Resources References Abudayyeh, O. O., Gootenberg, J. ...probably still edit genomes. Understanding the different types of CRISPR systems can be as confusing as...effector may have the same name but be completely different! They are merely linked by a similar function....Class 1 CRISPR systems use multi-Cas effectors, referred to as Cascade (CRISPR-associated complex for antiviral...compositions and mechanisms, with the biggest differences being the components present in the Cascade complex...like to explore Class 2 CRISPR in more depth, the Jennifer Doudna lab maintains an encyclopedia of Class ...

...duration of the conference (be warned, the hand sanitizer does go quickly!)  The Conference How many Addgenies... to brush up on your conference skills, check out some of our helpful conference blog posts before you...Scientific Conference Poster Session TipsHow to Make Friends and Meet People at a Scientific Conference New ...Getting ready for Society for Neuroscience Conference 2023? So are we! Come say hi to us at Addgene's...favorite content strategist, Jo. While we've all got different areas of expertise, we are each equally excited...invite you to join us! We'll also be sharing our conference experience on X, Instagram, and Bluesky.   Finally...

...subunit S12 of 30S ribosome Confers resistance to streptomycin Tn10   Confers resistance to tetracycline...coli bacteria. It’s hard to count the number of different commercial strains of E. coli currently available...the intestinal tract of animals. There are many different naturally occurring strains of E. coli, some of.... It was likely isolated in 1918 but was first referred to as “B strain” in 1942. The BL21 strain and ...F Host does (F') or does not (F-) contain the fertility plasmid. A low copy-number plasmid, encodes proteins...unless indicated otherwise fhuA (formerly tonA) ferric hydroxamate uptake, iron uptake receptor mutation...galactose only gyrA, gyrA96 DNA gyrase mutation Confers resistance to nalidixic acid  hsdRMS hsdR(rk-...

...performance under different imaging conditions (e.g. different emission filters) or different set-ups. Many...iii) aggregation tendency. Brightness Brightness refers to the fluorescence intensity of a fluorescent ...uptake by cells. To correct for this variation, a reference fluorescent protein that is expressed at an equal...be used. An assay developed in our lab uses a reference protein that is translated from the same open-...between the fluorescent protein of interest and the reference protein (Goedhart et al, 2011) allows for a precise... a set of fluorescent proteins (with the same reference protein) under identical conditions, a ranking...for co-expressing mTurquoise2 with mVenus as a reference protein is available from Addgene. Another way...

...for Neuroscience 2017 conference in Washington DC (#SfN17). This massive conference attracted over 30,000...-depth exposure to neuroscience prior to this conference was a rotation in Qiao Zhou’s lab in graduate...school where I worked on a project aiming to dedifferentiate mouse brain cells into neural stem cells. I...biologists. This was not at all the case. The conference featured people who work in sociology, behavior... Collider and which featured a talk about the different parts of the brain stimulated by stories. In retrospect...neuroscientists are using techniques from across different disciplines to better understand neurobiology ...develop technologies that will have impacts on many different facets of life. It also seems as if many of the...

...unaccounted for, may lead to different results. Different setting, different population, new genetic variation...obtaining a different result after conducting a replication study. An important difference between the...images, measuring the main outcomes in several different ways, and other questionable research practices..., simple noise in the process of sampling and inference-making. Even perfect transparent and rigorous ...variation, or different handling can all be important.  If we eliminate the problems above: unclear methods...previously unknown variable actually makes a big difference for the robustness of the findings.   How do...investigated to look for unexpected trends or differences between groups. Hypothesis generation is critical...

... activate different G proteins, work in different neurons and neuroglia, and have different effects on...Addgene’s chemogenetics guide.     References and resources References Kroeze WK, Sassano MF, Huang X-P,...These receptors can be activated by an array of different ligands, including hormones, neurotransmitters...is estimated that there are approximately 950 different human genes encoding GPCRs (Takeda et al., 2002...receptor, which can then activate expression of a luciferase reporter in the nucleus. The Roth lab showed ...activation via bioluminescence resonance energy transfer 2 (BRET2). Check out our blog post on GPCR signaling...track the activation and internalization of two different receptors- angiotensin II type 1 receptor and ...

...discussed are compiled for reference below. Happy virus making!  References and Resources Kulanayake,...the delivered cargo (often a gene) is called the transfer plasmid while the other plasmids with packaging... or something similar, and is contained on the transfer plasmid. If expression is the goal, a promoter...promoter and terminator regions and are slightly different in sequence. Both LTRs do share common repeat ...Adeno-associated virus AAV’s elements are quite different from gamma-retrovirus, but they perform very similar...delivered and expressed in the host cell from the transfer plasmid. Viral packaging and entry The viral packaging...which allows for altered tropism of AAV to target different tissue or cell types. AAV also requires some back...

...everyone at this specific conference, and I was able to include and exclude different modules for this workshop...something about it. Building a diverse team with different talents and expertise Fortunately, the eLife ambassadors...realized that holding the next one at a scientific conference could potentially enable reproducibility practices...it physically to instruct the workshop at the conference, but because each contributor made the materials...open and modular nature of the workshop content: different parts could be mixed and matched to serve a specific...the lookout for upcoming workshops at future conferences this year. Workshops are confirmed for the Evolution... having a workshop organized at your upcoming conference or you’d like to join or contribute to this collaboration...

...antibody’s buffer is compatible with your conjugation kit, since many common antibody buffers and buffer additives...are provided in a buffer with the antimicrobial agent sodium azide, which can interfere with conjugation...good base from which to dive deeper!References and resources References Bendall SC, Nolan GP, Roederer M,...love FPbase’s Spectra Viewer tool for comparing different fluorophores. We also touched on fluorophores ...chemical coupling can be achieved through several different mechanisms. Most take advantage of chemical groups... resources on the Addgene blog Antibodies 101: Buffers, Storage, and ConjugatesAntibodies 101: The Basics...

...exact location of the TSS. Different databases annotate the TSS in different ways, and it has been shown...phenotype of interest. This blog post will discuss differences between these approaches, and provide updates...Doench et al., 2017, and Hanna et al., 2020) and references therein. Nom nom nom. Me love gRNAs! Comic...are so few gRNAs to choose from that sequence preferences must largely be ignored. Created with ...few potential gRNAs. While SpCas9, with a PAM preference of NGG, is still the most widely-used Cas enzyme...Cas12a enzymes, and engineered variants thereof offers additional PAM options that can greatly expand ... with BioRender.com. Two newer technologies offer an alternative to HDR for introducing edits. The...

...section, teams are assigned different groups of three genes selected from different parts of the Met pathway...first select a multi-gene pathway to study and a reference organism where deletion strains are available....Students next transform deletion strains from the reference organisms with the overexpression plasmids. Finally...species has a similar function to the gene in the reference species. We have been using yeast models (3, 4...yeast, Saccharomyces. cerevisiae, makes a great reference organism for the project because it is non-pathogenic.... Over time, the two genomes have become very different due to a whole genome duplication and extensive...cloning to insert the coding sequences for ten different MET/Met genes (Figure 1) from the two yeast species...

... Types of ELISA There are four different types of ELISAs, which differ in the way the antigen is detected... to run an ELISA in your lab!   References and resources References Boguszewska, K., Szewczuk, M., Urbaniak...Antibodies are used in many different experiments that require scientists to detect proteins in their...antibody and the detection antibody recognize different epitopes on the target protein, so they do not... for which multiple antibodies that recognize different epitopes are available; this type of ELISA is ... are coated with another protein — known as a reference antigen — that can also bind to that antibody....antigen as the other types of ELISAs require two different antibodies. As mentioned above, it can also be...

...reads are mapped against a reference sequence, if one was provided. If no reference is given, de novo assembly...functioning properly or be something completely different than what was originally intended. Sharing unverified...scientists worldwide. Depositing scientists provide different types of data for each of their plasmids, from...plasmid sequence de novo (without the use of a reference sequence) using a bioinformatics pipeline developed...scientists to easily visualize and identify sequence differences between our NGS results and deposited sequences...mismatches in significant plasmid elements. Which differences are considered a QC issue is reviewed yearly ...approaches to confirm plasmid sequences without a reference; and identifying mixed plasmid and nucleotide ...

...drawbacks of different methods (for comprehensive protocols for all of the methods discussed here refer to Kutner...the correct transfer plasmid was used in the transfection; if preparing several different lentiviruses...assaying for expression of a gene carried on the transfer plasmid or quantifying the number of viral copies...resistance-genes, or the transgene itself. Many researchers prefer to design primers that target the common features...utilizes the antibiotic resistance genes carried on transfer plasmids. Target cells are transduced with serial...that standard conditions be used when comparing different batches of virus. In addition, some viral preps...step of a successful transduction experiment. References 1. Kutner, Robert H., Xian-Yang Zhang, and Jakob...