...  7 ways to strengthen your new relationships Here are some ways to go about strengthening a new relationship...events for one organization or go to the same conferences/meetings over and over to get to know the regulars...often the easiest way to build relationships and strengthen them over time.  Things to avoid when following...starfish shape. You can ask me about the different arms of this starfish sometime, but as of yesterday... was only one other person in my class who had children. When I had my baby, we became closer. She supported...really understood what I was going through. (Parenthetically, when I called the lab to ask my benchmate ...come across your contacts over the years in many different ways. This leads me to my last tip: It’s usually...

...plasmid cloning, and viruses. References Baltrus, David A., and Karen Guillemin. "Multiple phases of ...unique to the lab. Let’s take a look at these different methods of DNA insertion. Transformation Transformation...Various factors promote natural transformation in different bacteria such as growth phase of the cells (Baltrus...plasmid, the genes on the plasmid can give them different phenotypes. For example, this plasmid encodes ...cell. Once inside, phages can follow one of two different life cycles: lytic or lysogenic. Lytic phages ...a common tool used by scientists to introduce different DNA sequences of interest into a bacterial cell...through cell division. You can find all kinds of different lentiviral and AAV plasmids as well as ready-to-use...

... activate different G proteins, work in different neurons and neuroglia, and have different effects on...internalization of two different receptors- angiotensin II type 1 receptor and the β2 adrenoceptor. GPCRs in chemogenetics...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...

...St Pierre is currently a graduate student at Northeastern School of Nursing and currently works as a registered...low-income communities to educate parents about resources that can help their children thrive academically. Rose...helps you find internships and scholarships in different subject areas!    2. Resume and cover letter...your relationships to the person as well as their current sphere of influence. In other words, the longer...the better, and their leadership status in their current field of expertise augments their authoritative... while you make the ultimate case: why you? 7. Strength in numbers Encourage your close peers to come ...C. St Pierre! Roodolph (Roo) P. St Pierre is currently a doctoral candidate in the Chemical Biology Program...

...international mentoring conference held at the University of Valencia (Spain). At this conference, mentors and mentees...international mentors. Universities from five different geographical locations in Spain were enrolled:...mentees held scheduled monthly progress video conferences and had real-time correspondence via e-mail throughout...Mentorship Program and the International Mentorship Conference In addition to a pre-program self-assessment ... exchanged impressions of the IMP experience. Conference highlights included remarks by the Spanish Secretary...community overseas. Official representatives from different Spanish universities and high level public and...their support and hope for the IMP program. The conference also honored IMP Mentors & Mentees with awards...

...NmCas9 and St1Cas9. Each ortholog was fused to a different fluorescent protein to create three distinct colors...orthologs is key: since each ortholog requires a different PAM sequence, a gRNA designed for one dCas9 should...specific for telomeric sequences and showed that different fluorescently labeled dCas9s are efficiently directed...target sequence. They succeeded in labeling two different pairs of chromosomes using gRNAs specific to sequences...specified by PAM sequence, each color requires a different Cas9 ortholog, and the target sequence must be... green, and red. In contrast, a gRNA with two different hairpins will produce a secondary color cyan, ...sequence (as noted in the table). gRNAs with two different hairpin sequences produce yellow, cyan, or magenta...

...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...working with communities, not for them - Another strength of this collaboration is that it is driven by ...the lookout for upcoming workshops at future conferences this year. Workshops are confirmed for the Evolution...

... words, they were leaving these meetings with different interpretations of what we discussed and decided...scientists learn how to communicate their work differently based on their audience. How you talk about your...at a poster is going to vary depending on the conference you’re attending and who approaches you. You ...with professionals from other backgrounds at conferences or networking events. And if you’re a wet lab...fellow team member. But, at Addgene, we use seven different types of “plates” (Fig. 1). When you have to tell...certainly wasn’t the only problematic word. How many different things do you simply refer to as a “tube”? We... they could in fact be interpreted in several different ways depending on the team. When Tony, our corporate...

... Evaluating Gene function by RNA interference (RNAi) RNA interference (RNAi) is a powerful tool for performing...genes. Double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) to knockdown expression of specific gene...morgani, and Phagocata gracillis. There are 11 different genes cloned into plasmids to investigate for ...WISH technique. The expression patterns of 11 different planarian genes (all of which are available through...phenotypes of RNAi knockdown treatment should be apparent after three dsRNA feedings. Some of the phenotypes...and if you make new plasmids for investigating different planarian genes, we encourage you to deposit them...researchers can easily investigate in planarians. References 1. Alice Accorsi, Monique M. Williams, Eric J...

... 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...enzymatic reaction. Types of ELISA There are four different types of ELISAs, which differ in the way the antigen...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...

...you can quickly test that part in different contexts and plug different characterized parts into a TU! What...works on genetic code expansion.  References and Resources References Weber E, Engler C, Gruetzner R, Werner... then chances are you have been in the cloning trenches yourself. Modular cloning, or MoClo, is a systematic...be assembled into many combinations. Even with current technologies, it can be extremely hard to predict...nature of MoClo allows users to test a variety of different orientations and components within the system ... a great tool to deliver many combinations of different candidate genes at once to identify the best cocktail...and gene editing that use MoClo, for which many different MoClo kits have already been established. Keep...

...collaborate with specialists in different fields that can provide different insight into an area - not just...you are looking to move to a different university, perhaps in a different country, choose where you go...collaborators It’s a good idea to move around to different labs during your career; you learn new techniques...I hope to discover. This section may make the difference in whether you get funding or not; applications... grants too There are actually quite a lot of different pots of money available for researchers, so look...gained in writing small travel grants to attend conferences and a seed funding grant for a proof-of-principal...mechanisms of cell division. Recently Seán got the entrepreneurial bug and has set up an ELISA kit company on ...

...concentrating a viral stock. Different titering methods can reveal differences in physical versus functional...negatively-charged membranes (Davis et al., 2002). Polybrene is one such cationic reagent that is commonly ...efficiency by 10-fold (Davis et al., 2002). However, polybrene is highly sensitive to freeze-thaw cycles and ...be stored in single-use aliquots. Never use a polybrene solution that has been thawed more than once. ...in cells that are sensitive to the toxicity of polybrene (e.g., hematopoietic or primary cells), and has...packaging cells for infection If your transductions aren’t working, you can check if virus is being generated...up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate...

...) signal. While many studies focus on promoter strength as a determinant of gene expression levels, the...these will not be detailed here, but additional references are provided at the end.  Nearly all common bacterial..., and III), each responsible for transcribing different types of RNA: Polymerase I is responsible for ...that each eukaryotic RNA polymerase terminates differently. Polymerase III, for example, relies on a specific...Rho-independent termination found in prokaryotes. This is different than Polymerases I and II, which both require ...factor dependent, Polymerases I and II employ different mechanisms to terminate transcription. Polymerase...and/or use weaker poly(A) signals like BGH.  References Peters JM, Vangeloff AD, Landick R. Bacterial...

... transposition.  Target site: Different transposons insert at different DNA sequences or target sites....specific locations in the genome.   References and Resources References Feschotte C, Pritham EJ (2007) DNA... inserted into new target sites. Created in BioRender.com.   Class II TEs: DNA transposons Class ...a target site duplication (TSD). Created in BioRender.com.   Autonomous vs. non-autonomous TEs You...commonly used in the lab While there are many different types of transposons, DNA transposons are most... vectors.   Let’s take a closer look at three different transposon systems adapted for use as research...(Sandoval-Villegas et al., 2021). Target site preference and host range are important to consider when...

...them. For example, gene knockdown with CRISPR interference (CRISPRi) has been limited to lab-adapted bacteria.... Mobile-CRISPRi from the labs of Jason Peters, Oren Rosenberg, and Carol Gross provides a streamlined...plasmids for transfer to recipient cells during a tri-parental mating scheme. Mobile-CRISPRi integrates downstream...following IPTG induction of conjugation genes and bi-parental mating. Following excision from the B. subtilis...a recipient bacteria strain of interest by tri-parental mating. Donor bacteria contain a chromosomal copy...components is transferred to recipient bacteria by bi-parental mating. The ICE integrates into the trnS-leu2 ... of guides, promoters, selection markers, and different versions of dCas9. Following integration, Mobile-CRISPRi...

...subtypes of A–G, which come from different species and will have different compositions of Cascade. Although...through G, with different compositions of the Cascade complex. Created with BioRender.com.   The Cascade...degradative power in the future! References and Resources References Loeff, L., Brouns, S. J., & Joo, ... archaea. Class 1 is further broken down into different types, with Cas3 systems making up type I, the... complex from CRISPR type I-E. Created with BioRender.com. Thanks to this complex, developing Cas3...of the Cascade-Cas3 mechanism. Created with BioRender.com.   The reeling mechanism The reeling mechanism...

...discomfort of the mice, and restraint method. Many different substances can be injected, including cells, drugs...injection routes in mice. Image created with biorender.com.   Needle techniques Intravenous Intravenous.... They are a popular choice for administering different drugs and viral vectors. Like IV injections, intraperitoneal...sciatic nerve. Mice likely experience some muscle soreness after injection, like you might after getting ...as both procedures target the lungs. The main differences are that intranasal is easier, less invasive,...be flexible or metal, depending on researcher preferences. It sounds complicated but has a pretty low learning... to 10 mL/kg Intermediate Resources and references Additional resources on the Addgene blog Mouse...

...leverages the strengths of PRIDICT/PRIDICT2.0 while accounting for some of the inherent inaccuracies of...focus on gene editing techniques. References and resources References Akhtar, W., De Jong, J., Pindyurin...means we could design hundreds to thousands of different pegRNAs for each desired edit, it simply becomes...many more. Once we had our datasets with all the different features and editing efficiencies, we could train...offline via GitHub. The model provides users with different designs of pegRNAs, ranked by their PRIDICT/PRIDICT2.0... predictive power is sometimes limited across different targets due to variations in chromatin context... Our recent studies (Mathis et al., 2024) and concurrent research by the Shendure lab (Li et al., 2024...

...overlapping markers. Compensation/reference control(s) Compensation or “reference” controls are samples stained...spectral flow cytometry are more alike than they are different, there are a few aspects in which spectral flow...complex mathematical algorithm and single stained reference controls to distinguish one fluorophore from another...   Figure 3: Similarity index across 12 different colors on a three laser spectral flow cytometer...conventional or spectral flow. In spectral flow, reference controls tell the software to look for that exact...is not the same as your single color control. Reference controls: Beads or cells? Compensation beads are...new panel on a spectral cytometer, you prepare reference controls of both beads and cells for a single ...