... 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...

...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...

...created using BioRender.com (Link opens in a new window) . Addgene Website & Blog References Website CRISPR..., A., Herreno-Pachón, A., Benincore-Flórez, E., Karunathilaka, A., & Tomatsu, S. (2024). Current strategies...Alternatives Resources Plan Your Experiment Web References PAM Sequences Glossary Publications CRISPR Overview...mutation within the targeted gene. However, the strength of the knockout phenotype for a given mutant cell... into high fidelity Cas9s (hfCas9) . There is currently no defined measure of fidelity, so sometimes they...pyrimidine-to-pyrimidine) Base transversions - edit to different classification of nucleotide base (purine-to-pyrimidine...genetic changes, including moving entire genes to different locations, by combining other gene editing mechanisms...

...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...

...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...

...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...

... 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...

...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...

...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 ...

...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...