We narrowed to 841 results for: RING
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TypeBlog PostPublishedFeb. 11, 2025, 2:15 p.m....more gRNAs must be used along the same chromosome. During DNA repair, there is a chance that the two breakpoints...Harnessing type I CRISPR–Cas systems for genome engineering in human cells. Nature Biotechnology, 37(12),...CRISPR-Cas Systems INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons Additional resources...
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Academic vs. Industry Postdocs
TypeBlog PostPublishedNov. 13, 2018, 1:22 p.m....an industry postdoc is something that you’re considering. Like anything, there are pros and cons to postdocs...number of steps, decisions and people involved is staggering. The process is complicated. If you’re interested...all about teamwork. A single individual could not bring a drug to the market, so people in industry have... -
Plasmids 101: Optimizing Plasmid Yields
TypeBlog PostPublishedMay 26, 2016, 2:30 p.m....media with supplements such as magnesium salts, buffering agents, and/or supplying additional carbon sources... yield. Start with a single, fresh colony Sub-culturing directly from a frozen glycerol stock or agar ...harvesting and miniprepping at various time points. Measuring the optical density of the culture can indicate... -
Tips for Using BLAST to Verify Plasmids
TypeBlog PostPublishedMay 29, 2014, 1:29 p.m....blastn program uses a smaller word size and lower scoring penalties for mismatches and gaps in the alignment...blastp algorithm parameters. We use the default scoring matrix BLOSUM62, but you may want to check the ... multiple protein sequences, particularly for comparing different isoforms. In addition to our favorites... -
Antibodies 101: Producing Recombinant Antibodies
TypeBlog PostPublishedAug. 13, 2024, 1:15 p.m.... It may be helpful to incorporate regular feeds during this incubation time to maintain cell viability...consider increasing the number of washes performed during your harvest. Validation Recombinant antibodies...temperature shifts. Of course, we are also cautious during the aliquoting and storage process to prevent dropped... -
Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!
TypeBlog PostPublishedDec. 3, 2019, 11:38 a.m....for distribution. If your experiment involves delivering a calcium biosensor using AAV, you might decide...viral preps instead of plasmids? The available filtering options now change dynamically based on the type...target organism. Clicking through to RNA Editing brings us to a page with a basic overview and figure, ... -
An Introduction to Adenovirus
TypeBlog PostPublishedOct. 3, 2023, 1:15 p.m....inhibition of the innate immune system; and reconfiguring of the nuclear compartment, gene expression,...’ functions are rescued by helper plasmids used during production and/or by genes present in the HEK293T...license. The promise of adenoviral vectors This brings us to the final topic, which is that natural AdVs... -
Lentiviral Vector Uses and Overview
TypeBlog PostPublishedMay 19, 2016, 2:30 p.m.... the genome, they could promote oncogenesis by altering local gene expression. Self-inactivating (SIN)... PubMed Central PMCID: PMC4486245. 5. Patient monitoring and follow-up in lentiviral clinical trials. ... Mary Gearing... -
Plasmids 101: Colony PCR
TypeBlog PostPublishedMay 12, 2016, 2:30 p.m....up colony PCR reactions is nearly identical to preparing a standard PCR reaction: combine template, primers...other colony PCR tips are highlighted below. Preparing template: Pick a single colony with a sterile ...to the PCR reaction. The bacteria will be lysed during the initial heating step of the PCR reaction. A... -
Cpf1: A New Tool for CRISPR Genome Editing
TypeBlog PostPublishedOct. 14, 2015, 2:30 p.m....requirements. Cpf1’s preferred PAM is 5’-TTN, differing from that of Cas9 (3’-NGG) in both genomic location...repair templates should remove the PAM sequence. Comparing Cpf1 and Cas9 on-target and off-target efficiency... Mary Gearing... -
A Career in Grants: How To Become a Grant Professional
TypeBlog PostPublishedAug. 20, 2015, 2:30 p.m.... training, mentoring and coaching services, including Smartegrants, to help those entering the Grants ...Twitter Chat, and then our grant training and mentoring. How do I become a grant professional? Employment... -
Neuronal labeling with Spaghetti Monster
TypeBlog PostPublishedAug. 14, 2018, 12:42 p.m....proteins (FP) have also been important in mapping, monitoring, and manipulating neuronal circuits. Epitope ...tags, low FP expression may be insufficient for monitoring a weakly expressed POI. Spaghetti Monster - A...fluorescent protein structure and color variants." Cold Spring Harbor Protocols 2009.12 (2009): pdb-top63. PubMed... -
The PAM Requirement and Expanding CRISPR Beyond SpCas9
TypeBlog PostPublishedAug. 20, 2020, 2:30 p.m....first non-Cas9 CRISPR protein adapted for genome engineering was Cas12a (formerly Cpf1), a nuclease that generates...comment below. Note: Jennifer Tsang and Mary Gearing contributed to the update of this post. References... genome engineering... -
Finding nucleic acids with SHERLOCK and DETECTR
TypeBlog PostPublishedApril 16, 2020, 1 p.m.... the promiscuous cleavage and degradation of neighboring ssRNA and ssDNA by Cas13 and Cas12a, respectively....org/10.1038/s41587-020-0513-4 Chen JS, Ma E, Harrington LB, Da Costa M, Tian X, Palefsky JM, Doudna JA...Essletzbichler P, Dy AJ, Joung J, Verdine V, Donghia N, Daringer NM, Freije CA, Myhrvold C, Bhattacharyya RP, Livny... -
Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)
TypeBlog PostPublishedJuly 8, 2025, 1:15 p.m....success depends directly on the choices you make during the early stages of the process. Some questions...which acceptor vectors and parts you will be using during the different levels of your MoClo assembly. Know...fragment can be used directly in your assemblies. Preparing reagents Keep it cool You’ve done all the learning... -
Antibodies 101: Reading a Flow Plot
TypeBlog PostPublishedFeb. 6, 2024, 3:13 p.m....subpopulations of cells, you’ll soon find yourself encountering flow data in the literature. Data reported from...of flow cytometry is to use increasingly more stringent markers to identify your specific cell population...are used to separate single cells from doublets, ignoring anything not within the gate of the first plot... -
Antibiotic Resistance: An Old Solution but a New Problem
TypeBlog PostPublishedNov. 10, 2016, 3:30 p.m.... resistance can be the result of one bacterium sharing genes with another, an accidental transmission ...means to defend themselves from these naturally occurring antibiotics. When we use antibiotics to treat ...prescribed to them This may seem counter-intuitive considering the other three scenarios, but a short treatment... -
The Challenges of Cell Culture
TypeBlog PostPublishedMay 5, 2016, 2:30 p.m....majority of cell line problems come from people sharing cells: the black market of cell biology. Engaging...cell culture, microbiology, or virology without acquiring materials from this bank. I am often asked, “What...cells to be used and STR profiling to be conducted during projects to ensure the results are reliable. With... -
Important Considerations in Optogenetics Behavioral Experiments
TypeBlog PostPublishedOct. 1, 2015, 2:30 p.m....successful inhibition of hippocampal neurons. Initial monitoring Second, this may sound rudimentary, but before...exciting—and unanticipated—road of discovery. Monitoring behavioral output The appropriate behavioral ...science writing. Derek is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org... -
Tips for Using FRET in Your Experiments
TypeBlog PostPublishedNov. 5, 2014, 3:41 p.m....The first time I heard about FRET during a journal club, my guitarist brain automatically thought about... recently and is the most rigorous method for measuring FRET. FLIM measures the fluorescence decay time...MW. Fluorescent proteins for FRET microscopy: monitoring protein interactions in living cells. Bioessays...