...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...

... 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...

...yours does and then use that time! Different platforms have different quirks, so remember to practice on...that requires its students to do rotations in different labs before selecting an advisor, it is always...points and keeping them at-hand, you can quickly reference your questions about the PI’s research or the ...the name of the award you won for your conference presentation. It can help soothe your nerves and make ...promoted during COVID-19, online meetings with current students, faculty, and staff have become a new ...of just the program requirements: by talking to current students about the work environment and social ...using the virtual world to your advantage can strengthen your application and help you decide if a program...

...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 ...

...Organizing a science conference for the public How does one organize a public scientific conference? As a member...first-time event called DayCon. DayCon is a one-day conference aimed at the general public that provides an ... commitment of SITN members. SITN and DayCon Currently in its 16th year, Science in the News has two ...scientists and the public and help make science more transparent. Second, SITN helps graduate students develop...graduate student at Harvard, who wanted to provide a conference-style experience that was accessible to the general...picked a theme (Bio-Medley) to showcase SITN’s strength in the biomedical sciences and decided to offer... make DayCon a yearly event and to expand the conference to accommodate a larger number of attendees. ...

... of antibiotics, but the field is currently undergoing a renaissance. Yosef et al. designed a two-phage...lysogenic phage, rendering the population sensitive to antibiotics. Created with BioRender.com. Phages ...swift, as few as five years after introduction. Current antibiotic strategies are nonspecific - they harm...defense against bacteria. Created with BioRender.com.   The problem of antibiotic resistance ...alter an individual’s native, beneficial flora. Currently, there is no antibiotic strategy available to ...mixed population, represented here by cells of different colors, specifically targets bacteria with the...ground in the fight against bacterial superbugs. References 1. Citorik, Robert J., Mimee, Mark, and Timothy...

...the host. The cause, manifestation, diagnosis, recurrence, and treatment of cancer have been extensively...compromised either due to age, infections, or other strenuous biological changes. In contrast to the acute immune... cytokines like TGFβ and IL-6. However, when strengthened through efforts to boost the activity of the... most commonly used immunotherapeutic agents. Currently the FDA has 12 approved monoclonal antibodies .... Sipuleucel-T by the Dendreon Corporation is currently the only FDA-approved DC therapy for prostate ...only just touched the surface of immunotherapies currently under study. Check out this review for an in-depth...manifestation to their specific interactions with different cell types. A thorough understanding of the genotypic...

...soils. Dendrobatidae (Lauren O’Connell)  Poison frogs are great for studying parenting strategies and make...land. Current tools are all for Xenopus (aquatic frogs), but there are few genetic tools currently for ...Genomic Tools) Program and I met scientists from 21 different labs who clearly also share this wonder of the...convergent evolution because electric organs arose in different branches of the evolutionary tree at least 6-7...editing in the brain allows the squid to create a different protein. Did you know that most cephalopods die...behavior. For example, two similar species have very different social behavior. One is monogamous and one is ...new tools being developed and shared for many different organisms. Do you have a favorite organism you...

...for other antibody topics.     References and resources References:  Björck L, Kronvall G (1984) Purification...elute and measure the target. However, the two different approaches allow for optimized strategies for ...amounts of target proteins or antibody bonding strengths.    . Steps of an IP: 1) Protein extracts... appropriate for most situations.  A slightly different method is desirable if the antibody is expected...Protein G have an affinity for immunoglobulins, the strength of the interaction varies between antibody host...than those of agarose beads, despite the size difference. Magnetic beads are particularly advantageous...many factors, including salt concentration, ionic strength and pH, affect binding the lysis buffer may need...

...host species. Created with BioRender.com. How are shuttle vectors different from other plasmids? If ...protects against any of them. But different aminoglycosides work on different host species: kanamycin targets... unrelated host species, even from completely different kingdoms of life. That’s convenient for researchers... Shuttle vectors have to address several big differences in the ways vectors are used across species: ...resistance gene on your vector might require two different selection molecules! It all depends on how the...Auxotrophic selection Another option is to use a different method of selection entirely. Auxotrophic selection...host range,” these terms are typically used in different (though overlapping) ways. Broad host range plasmids...

... can separate the DNA strands. Created with BioRender.com. Once DnaA breaks the local hydrogen bonds...Prokaryotic chromosomal replication. Adapted from BioRender template. Since DnaA binding is necessary ...proteins also contribute to this regulation in different ways: methylating newly synthesized DNA, blocking...numbers can allow RepA to link iterons from two different plasmids, forming a pair of “handcuffs” that blocks...RepA proteins form complexes between iterons on different plasmids in a process called "handcuffing," which...folding, blocking replication. Created with BioRender.com. Plasmids under stringent control of replication...was updated by Emily P. Bentley in June 2024.  References and resources Further Reading Quiñones-Valles...

...expect, different companies produce different products and therefore need people with different areas of...: vast practical lab expertise, experience in different scientific fields, and knowledge of how to troubleshoot... as: “Can I use this product under conditions different from those specified on its data sheet?” or “Do... questions customers ask, and there are also differences in customers’ personalities and their expectations...do jobs in technical support have? There are different names for technical support roles. Examples are...distinction between these job titles, they may be different regarding the levels of lab experience and scientific...communication skills by working with people from many different backgrounds and realizing that the same approach...

...virus:  Lowering cytotoxicity. Different envelope proteins have different levels of cytotoxicity and the...Altering sensitivity to serum. Different pseudotyped viruses are differentially inactivated by nonspecific...Table 1 (Cronin et al., 2005)     References and Resources References: Crawford KHD, Eguia R, Dingens AS...protocol for generating lentivirus, but you use a different envelope glycoprotein instead of the wild-type...review articles to find tables and information on different viral membranes, tropism, and advantages and disadvantages....084 Li Q, Liu Q, Huang W, Li X, Wang Y (2017) Current status on the development of pseudoviruses for ...

...going to cite hundreds of references) and it’s better to keep your references organized from the beginning...the thought of writing a review, or if you’re currently stuck trying to write one, hopefully this post...criteria to shape the document. Get and use a reference management program (e.g., EndNote, Papers, Mendeley...Mendeley, etc.) You’re going to be managing a lot of references. I cite as I write, meaning I use the software...in real time as I wrote, so each statement was referenced even in the roughest version of the draft and...that people can go back and read the original reference for the data. For example, we summarized how frequently...place to get a sense of the similarities and differences among proteins in the class. Offer your perspective...

...educational training, and conferences, and synthesizing this knowledge into a coherent narrative that gives...best with diverse teams, to leverage the preferences of different personality types, and to devise strategies...biotech clients. In this realm, there are a few different kinds of challenges that consultants get called...companies. Medical Affairs groups work on building awareness among physicians and scientists about clinical...consulting firms coach and counsel employees on the different personality and work styles (e.g., Myers-Briggs...prepare you for a successful career in several different fields. While some people continue to enjoy the...BioCentury can be an engaging way to learn about the trends and workings of the healthcare industry. Finally...

...doping to forensics, DNA sequencing to drug testing. You can find Nick on LinkedIn. References 1. Skloot...Expression Attenuates Proliferation and Invasion in Renal Cancer Cells, Marker-independent identification ...the cell lines deposited with them and used by different researchers are free of contamination and match...from this bank. I am often asked, “What is the difference between materials available from commercial repositories...the same cell line from a colleague?”. The main difference is the QC and validation carried out by the commercial... The same test is so good that it is used as a forensic technique (aka DNA fingerprinting) that can place... produce a new batch once you’ve passaged your current batch too many times or you suspect there is contamination...

...are communicating across disciplines or doing different tasks. But these challenges can be exaggerated...equal opportunity to contribute. Using video conferencing tools like Zoom or Google Hangouts is a great...great way to include researchers from different locations. Having virtual face-to-face meetings as opposed... collaboration includes scientists working in different time zones, it’s important that everyone can access...project at the International Cancer Immunotherapy Conference in Frankfurt, Germany When funding and circumstances...techniques are being performed the same way at different sites. I did a lab exchange as part of the collaboration...allowed me to investigate my protein of interest in a French clinical cohort, neither of which I would have ...

.... The plasmids within it contain a variety of different promoters followed by leader sequences (encoding... at the BIOFAB showed that expression of many different genes was predictable based solely on the promoters...BIOFAB Plasmid Set contain terminators of varying strengths centered within GFP/RFP bicistronic reporters ...or protease sites to remove the epitope tags. Different plasmids in the kit can be used to purify proteins...prokaryotic expression in recent years, with many different systems now available. However, it can be difficult...ORF clones to allow testing and comparison of different cell-free expression systems; the Cell-free expression...bind specific RNA bases. By swapping out these different modular repeat segments, you can create a PUF ...

...what we need to know in order to even assess the current state of research became more clear throughout ...that, even though postdoc associations across different universities are trying to compile more comprehensive...path,” but individual institutions can have many different names for the positions commonly thought of as...the Starting Salary for Postdocs to $50,000 (the current median postdoc starting salary as reported by Paula...Student Career Outcomes University websites do not currently publish career outcomes for PhDs or postdocs. ...achieved. Of note, it was highlighted during the conference that just increasing funding for academic research... At Addgene we try to do our part in boosting awareness of various career opportunities through our career...

...microscopy and beautiful imaging.   References and Resources References  He, Y., Zhang, T., Sun, H. et al...killing the untransformed seedlings or callus (undifferentiated plant cell culture), which is great for a ...desired plants! PCR or fluorescent markers are currently used in these situations to select against CRISPR...such as cacti and beets. Betanin belongs to a different chemical family than the red/purple plant pigments...it for selecting transformation events and am currently planning to use it to simplify my work with an...(Bottom panels) RUBY expression can vary from different transformation events likely due to insertional...lower expressing RUBY plant is a pale pink in transparent tissues, or in green tissues is slightly darker...