We narrowed to 527 results for: HA tag
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TypeBlog Post...variant has increased signal-to-noise ratio and better ability to record subthreshold voltage fluctuations...optical pooled CRISPR screening, Paul Blainey’s lab has developed a multiplexing approach to Cas9-based pooled... stabilize the 3’ end of the pegRNA. As research has just begun with PE7, it will be exciting to see future...limitations of traditional AAVs, the Becirovic Lab has developed REVeRT, for gene delivery using reconstitution...Voltron2 voltage sensors By Brian O’Neill New viral vector preps expressing the Voltron2 voltage indicator...-2 research New viral vector preps of Voltron2 voltage sensors AAV vectors for targeting microglia The...earlier Voltron indicators (a fusion of Ace2N and HaloTag, Figure 4), they determined that an A122D mutation...
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Hot Plasmids and Viral Preps - July 2021
TypeBlog Post...require heavy optimization. Claudia Vickers’s lab has designed novel SL fluorescent biosensors by integrating...showed that the receptor-based fluorescent biosensor has a high sensitivity (50 nM-500nM range) and a high...recent months: Our selection of GCaMP8 AAV preps has expanded. Check out the new selection of ready-to-use...performing genetics studies in organoids. The Organoid EasyTag System is a straightforward workflow for doing ... Figure 1: Example schematic from the Organoid EasyTag system, showing the workflow for inducible CRISPR... Image from Sun et al. 2021. The Organoid EasyTag workflow combines thoughtfully designed plasmid... -
The Strength of Story Telling
TypeBlog Post...Show that as the story has progressed, you or your lab or science or your CV has grown as well. Find your...Task, Action, Result) interview technique, which has been addressed in much better pieces than I’m able... your structure. Develop your protagonist In most cases, the protagonist will be you. Make sure to highlight...enjoy, remember, and listen to narratives — take advantage of that in your work The part of my job I most... -
Some Like it Hot: Thermostable GeoCas9
TypeBlog Post...Kcleave (min-1). SpCas9 has maximum activity from 35-45 °C, while GeoCas9 has maximum activity from 50...the recent discovery of GeoCas9 by the Doudna lab has opened up the field of thermophilic bacteria to CRISPR...stability in human plasma The last trick GeoCas9 has up its sleeve is its increased longevity in human...stable when incubated in reaction buffer alone, but has limited activity when incubated in diluted human ...50-70 °C. The key advantage of GeoCas9 is that it functions at temperatures much warmer than SpCas9... -
RNA Interference in Plant Biology: New Tools for an Old Favorite
TypeBlog Post...It is fair to say that CRISPR has revolutionized molecular biology and has enabled functional studies in...enzyme named Dicer (Figure 1). The plant kingdom has expanded its repertoire of Dicer enzymes and these...knockout methods of the same gene. However, recent work has called this prevalent assumption into question. Specifically...multiple target sequences are concatenated. hpRNA has been extensively used in plant research and many ...constructs are available to the community. Each construct has unique features, such as inducible expression of ...tool requires careful balancing of the inherent advantages and limitations of any technique in the context...interest. Tools for RNAi Arguably the greatest advantage of RNAi is the straightforward nature of the technique—all... -
Early Career Researcher Toolbox: Social Media for Scientists
TypeBlog Post...ElizabethKenkel) December 5, 2018 How social media has helped me in science It may have taken a bit of FOMO...Here are three examples of how using social media has helped me or my lab mates: For me, using social ...with social media as a scientist Using social media has enriched my professional development, and while it...helped you fit social media into your busy schedule. Has social media helped you advance your research or ...handle, but I wasn’t using them to my professional advantage. It wasn’t until I wrote a blog post about a research...start using social media to their professional advantage, I’ve selected five resources that have tips for... -
Troubleshooting Your Plasmid Cloning Experiment
TypeBlog Post...Service since 2009. He has developed a proprietary, low cost cloning protocol that he has used for cloning ...least 24 bp with the template and if the sequence has high GC or AT content, increase the overlapping region... most of the troubleshooting regarding this step has to be strategy specific, there are few general parameters...ratio of 1:2 (plasmid:insert), but this parameter has to be optimized according to the strategy adopted...satellite colonies and they form when all the antibiotic has been used. Don’t worry too much, just pick the big...many softwares out there than can help you at this stage and that can be used to simulate in silico cloning... -
Which Fluorescent Protein Should I Use?
TypeBlog Post...set as 1. Some proteins are very dim (e.g. TagRFP657, which has a brightness of 0.1) and this should be ...Fluorescence is the emission of light by a substance that has absorbed light. The emitted light is at a longer ...protein Excitation & Emission (ex/em): Each FP has its unique ex/em peak. Therefore, choose FPs that...detect the emission. For example, if your microscope has only two lasers, at 488nm and 561nm, you will not...pHTomato (pH sensors), HyPer (H2O2 sensor), ArcLight (voltage sensor), and iGluSnFr (glutamate sensor). More ...can fold in <10min at 37°C, mCherry takes ~15min, TagRFP ~100min and DsRed ~10hours. Temperature: FPs maturation... -
Lentivirus Production
TypeProtocol...10 min and then recheck the pH to ensure that it has not drifted. Filter the solution through a 0.22 μm...L-alanyl-L-glutamine (or alternative stable glutamine such as glutaGRO, Corning 25-015-CI) Low serum medium such as Opti-MEM...L-alanyl-L-glutamine (or stable alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add ...add 55 mL of heat-inactivated FBS and 5 mL of glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots...transfection to determine what ratio gives the highest percentage of GFP positive cells. Refer to the table below... -
pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs
TypeBlog Post...reprogrammed less efficiently, no such correlation has been found between the age of the patients from whom...potentially promote tumorigenesis in vivo. The phenomenon has been recorded in past gene therapy trials and remains...reprogramming factors To date, the mRNA transfection method has not been used successfully to derive iPSCs from PBMCs...However, reprogramming of PBMCs by episomal vectors has yielded hiPSCs at a low efficiency (0.001-0.03%) ...efficiency: the pCXLE episomal vector toolkit It has been shown previously that suppression of TP53 enhances...hiPSCs than age-matched fibroblasts [3]. These advantages make PBMCs the ideal source of somatic cells ... This is to avoid the chances of insertional mutagenesis, which is the integration of transgenes into ... -
DAP arrays for multiplex gene editing
TypeBlog Post...Recently, Xue Sherry Gao’s lab at Rice University has developed DAP arrays for efficient multiplex base...process and release its own gRNAs from a single array, has been a top choice for multiplex gene knockout or ...proper functions to enable efficient base editing and has yet to be developed for prime editing, both and their..., which is conserved in all living organisms and has previously been used for multiplex Cas9-nuclease ...lengthy promoters, such as U6 or EF1α promoters, has limited the number of gRNAs that can be packaged ...CRISPR tools, without compromising their strategic advantages in gene editing, while keeping the system to ... -
Year of the Camelid: Antibody Style
TypeBlog Post...The UN General Assembly has declared 2024 the International Year of Camelids. The declaration is intended... antibody libraries to generate new sdAbs. There has also been some interesting work done developing transgenic...al., 2021. The single-domain antibody field has come a long way since the serendipitous discovery...reagents. But for a quick refresher, some of the advantages of sdAbs over conventional antibodies include... simple structure sdAbs in research These advantages have contributed to sdAbs becoming valuable tools... -
Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More
TypeBlog Post...positions. There has been a lot of engagement with the Athena SWAN Charter and it has really strengthened...their pathogens. As Patron explains it, her work has always been focused on gene transfer, from transgenes...MoClo) Kit. What does it include? Patron: Sainsbury has a mandate to share and educate and enable plant science...quantitative data for regulatory elements, which has not been available for plants. Addgene: Will the ...strengthened the network of female scientists. Twitter has been really good for that too. I’m more in touch with...s research network here. She talked about the advantages of talking to people and representing yourself... -
Colony Formation Titering Assay
TypeProtocol...mL) x dilution factor e.g., If the 1:100,000 well has 75 colonies, then there are 75 colonies /1.5 mL x...L-alanyl-L-glutamine (or alternative stable glutamine such as glutaGRO, Corning 25-015-CI) Heat-inactivated FBS Polybrene...L-alanyl-L-glutamine (or stable alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add ...55 mL of heat inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots... -
Viral Vectors 101: Chemogenetics
TypeBlog Post...tools. The advantages provided by chemogenetics are instead that it is noninvasive and has longer-lasting... disrupting normal brain function. Chemogenetics has become a popular tool in the field as it provides... agonist for muscarinic receptor derived DREADDs has been clozapine-N-oxide (CNO). However, concerns have...are already approved for clinical use. Varenicline has been further modified to create ultrapotent PSEMS...will promote hyperpolarization. PSAM ligands The advantage of PSAMs lies in their ability to be specifically... -
Help Us Name Blugene’s New Virus Friend
TypeBlog Post... favorite! Our mascot who has supported us over the past eight years now has a new friend to join them...taking name suggestions using this form or the hashtag #BlugeneVirusFriend on Twitter. As with Blugene... -
Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone
TypeBlog Post...never went away. While he has frequent bouts with Impostor Syndrome, Joe has found his people through ... match people within similar fields (ASM Microbe has eight tracks), something we didn’t have enough people...notebook paper, despite the fact that it was gray. He has worked in molecular and microbiology for the last...conference hashtag I had, however, started a Twitter account, and was following the meeting hashtag. I noticed...Favorable conditions - Expansion of networks The last stage is Exponential Growth, named after the growth phase... -
Addgene Receives NIH BRAIN Initiative Grant to Create Open-Access Recombinant Antibody Resource
TypeBlog Post...Davis School of Medicine on this grant. Dr. Trimmer has created thousands of high-quality antibodies/nanobodies...mission This is the first NIH grant that Addgene has received. Addgene prides itself on being financially...The timing is ideal as the NIH’s BRAIN Initiative has been funding laboratories, such as Dr. Trimmer’s ...antibodies are plasmid-based and offer numerous advantages, including more secure archiving, unambiguous...AK, Anderson S, Bladen CL, Jones B, Aldred SF, Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS... -
Plasmids 101: Restriction Cloning
TypeBlog Post...and insert a linear fragment of DNA (insert) that has been cut by compatible restriction enzymes. An enzyme...one found on the left side of the image below. It has a promoter (blue arrow) followed by the restriction...finished plasmid Once it looks like your ligation has worked, you will need to pick individual bacterial...purpose. Ideally, once you know that your plasmid has an appropriately sized insert, you should send it...plasmid for more details. Once your complete plasmid has been verified, you’re ready to get experimenting!...gel purification. In gel purification, you use a voltage difference across a gel matrix (usually agarose...positioned toward the cathode (- charge). When the voltage is applied across the gel, the DNA migrates toward... -
Finding nucleic acids with SHERLOCK and DETECTR
TypeBlog Post... but inadequate access to reagents and equipment has become a bottleneck. In the last few years, scientists...nucleic acids at the atomolar level. The Zhang lab has adapted natural RNase activity of the Cas13 protein...Gootenberg et al., 2017 and 2018). While the Doudna lab has used Cas12a’s non-specific ssDNA degradation to develop...field (Myhrvold et al., 2018). Now, the Zhang lab has shared a protocol for using SHERLOCK to detect SARS-CoV...at atomolar levels, within one hour. DETECTR thus has the ability to rapidly detect nucleic acids with ...diagnostic applications. The company Mammoth Biosciences has been started based on DETECTR technology with the...Biotechnology. Like the SHERLOCK detection system, DETECTR has not been approved for diagnostics yet, but is undergoing...