...In previous posts for our Plasmids 101 series, we examined a number of important plasmid elements – promoters...and the region of DNA between the loxP sites is reversed.   Deletion: If the sites face in the same direction... cassette) upstream of a gene of interest will prevent gene expression in the absence of Cre. In the presence...recombination event in a single system. The Saccharomyces cerevisiae FLP-FRT recombination system is another site-...

...fluorogen A split fluorescent reporter with rapid and reversible complementation. Tebo AG, Gautier A. Nat Commun...options. While the BiFC interaction is typically irreversible and requires some time for chromophore maturation...maturation, splitFAST displays rapid and reversible complementation. TagBiFC (split HaloTag) provides maximum...important for their interaction. You can read our previous blog post for more on BiFC principles and experimental...

...low cost and quick point-of-care diagnosis kits (reviewed in Tsang and LaManna, 2020). To see even more ...uses loop-mediated isothermal amplification after reverse transcription (RT-LAMP) and a colorimetric readout...completed in just 45 minutes. The team deposited the reverse transcriptase, protease, and Bst polymerase constructs... this toolkit include ones for expressing MMLV reverse transcriptase and Taq polymerase that can be used...

..., you will need to correct it). It’s easy for reviewers to spot when an application is rushed (whether...that you ask a PI or mentor (your supervisor/ a previous supervisor / someone else in the department) to... times and will have useful advice about what reviewers in your area are looking for. Remember though,...for grant success The two most important things reviewers will be looking for in your research proposal ...

...broken during cross-link reversal (not shown in diagram). Cross-links are reversed so that DNA fragments ...cells (~5x107). Together, these two requirements prevent the use of CAPTURE with primary cells or rare cell...multiplexed CAPTURE 2.0 largely replicated interactions previously identified with the original CAPTURE method. ...

...new managers and I had a “Eureka” moment while reviewing the results of my Myers-Briggs test:  my undergrads...motivation, but he didn’t want to spend hours reviewing papers to piece together the best protocol;  he...associate feedback with end-of-the-year performance review, but feedback should be given throughout the year...References Becoming the boss. Harvard Business Review, January 2007. Three Rules for Managing a Successful...

...then dropped to a range in which the forward and reverse primers are stable. At this temperature the primers...can also be used to quantitate RNA levels using reverse transcription-qPCR (RT-qPCR). The first step of...process requires RNA to be converted to cDNA using reverse transcription and the cDNA is subsequently quantified...template is amplified using several forward and reverse primer sets.  Multiple template PCR - multiple...

... are voltmeters for cells! In this blog we will review how these indicators work, their general applications...of the membrane, a concern with several of the previously described systems. Several adaptations of this...   Viral Vectors and Voltage Indicators As previously mentioned, cellular voltage is measured at the... Components of the types of voltage indicators reviewed.   References and Resources   References ...

...Cas13-based RNA editing system would likely be reversible and would avoid genomic off-targets or indels... RNA knockdown. In initial tests, Cas13b from Prevotella sep. P5-125 (PspCas13b) yielded 62.9% average...editing plasmids here! Future directions Building on previous RNA targeting work with Cas13, REPAIR is the first...edit RNA has multiple advantages, including reversibility and use in non-dividing cells. With the expanded...

...ambitious, that’s because it is. Zimmer, whose previous books focused primarily on fluorescent proteins...racism and sexism in science, problems with peer-reviewed publications and funding, bioethics in the age...unlikely to be surprised by many of Zimmer’s revelations. The issues he discusses have been making the...of his book today--the day you are reading this review--in the current period of rapid flux, the chapter...

... functional copy of your plasmid of interest, preventing this issue. The genes and their antibiotics   ...produces beta-lactamase to degrade amp and therefore prevent cell death. One of the cool things about amp is...susceptible to degradation in agar plates. This prevents satellite colonies from forming while effectively...expressing a protein that directly binds to zeocin, preventing it from binding DNA. Zeocin is an all-in-one ...

...within a CRISPR guide can prevent the guide from binding to the genome or prevent Cas9 from cutting, both...genome could affect their analysis. Their work revealed that cell lines from individuals of African descent...Perturbation Platform to modify the CRISpick algorithm to prevent guides from being designed that target regions ...

... the Wisconsin research community were to: (1) prevent vendor lock in by making it easy to get data out... to a selected Slack channel for discussion and review. It’s also possible to capture Slack conversations... and/or import and convert it into RSpace. The reverse flow is also supported, so it’s possible to re-...making research data available for public access, review, and query. Gates Open Research and Wellcome Open...

... limited to 0-5%. This blog post covers a brief review of different basic BE technologies and their adaptation...making an original C-G pair converted to T-A or reverted to the original C-G by base excision repair mediated... by introducing a point mutation (H193Y). The reversion of this mutation by an adenine base editor would...in rice. In Genome Engineering: The CRISPR-Cas Revolution, Cold Spring Harbor Laboratory (2018). Additional...

...we give people the opportunity to speak out and prevent groupthink? Here are some ideas that I have found...a member of our Business Development team would review it. This survey generated 26 ideas, over half of...mistakes that kill your productivity. Harvard Business Review. Retrieved January 14, 2022, from https://hbr.org... spur innovation on your team. Harvard Business Review. Retrieved January 14, 2022, from https://hbr.org...

...participant in the training. In this blog we will review the preparation and planning you can do to build...different way of doing something then you’ve done previously) you can bring those questions to your training...questions throughout training, while others prefer to review their notes, process what they’ve just learned,...Show up prepared and take training seriously, as reviewed above. Offer to help clean up and/or make necessary...

...containing the desired edit adjoined to the sgRNA, a reverse transcriptase, and a nCas9. While this system has... Fig. 2: Prime editor engaging target DNA. The reverse transcriptase polymerizes DNA onto the target strand...successfully in experiments and published in peer reviewed journals. If you need to generate a new sgRNA ...your experiment. Keep an eye out for our upcoming review of these options in our CRISPR 101 series!  References...

...correlated with codon bias in both E. coli and S. cerevisiae. Protein folding - If a protein is encoded by...Gene optimization mechanisms: A multi‐gene study reveals a high success rate of full‐length human proteins..."Evolutionary conservation of codon optimality reveals hidden signatures of cotranslational folding." ...26186290. PubMed Central PMCID: PMC4794256. This review provides a great overview of codon usage bias ...

... such challenges involves using knowledge from previous consulting experiences, educational training, ...productivity, and many have mechanisms in place to prevent the burden from getting overwhelming. These mechanisms...teams are facing a crunch, and using lessons from previous cases to better staff future teams. As with any...how much my scientific background, training in reviewing literature, and critical analyses skills were ...

...In a previous Plasmids 101 blog, we reviewed the salient features of several popular strains of E. coli...over-expressing a foreign protein in E. coli. We will review the potential pitfalls of recombinant protein expression... problem for toxic protein products, which can prevent the culture from reaching the desired density within...