We narrowed to 604 results for: abo.2
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TypeBlog Post...out for help so you can get the support you need. 2) Prepare an agenda for every meeting Supervisors are... I heard about supervisors in all of the above categories. In addition, I also learned about two other...Complainers, who spend most of their time moaning about their troubles, Silent or unresponsive clams, ... students and postdoctoral fellows are hesitant about expressing their ideas, especially if their supervisors...it is interfering with your work. Avoid talking about your emotions, such as frustration or anger, because... like the problem to be resolved You know more about your work than your supervisor does. You can save...Listen to your supervisor’s viewpoint and brainstorm about mutually beneficial solutions Your supervisor might...
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Lentivirus ddPCR Titration
TypeProtocol...Activation 95 10 2 1 Denaturation 94 0.5 2 40 Annealing/Extension 60 1 2 40 Enzyme Deactivation 98 10 2 1 Hold ...diploid cells, thus the reason for multiplying by 2. $$V = 2*{copies\ RRE \over copies\ RPP30}$$ Use the viruses...diluted 2-fold serially, the concentration of RRE positive droplets should decrease by a factor of 2 across...Lentivirus is generally considered biosafety level 2+. Please ensure that you are in compliance with your...channel pipette 200–1000 µL single channel pipette 2–50 µL multichannel pipette 20–200 µL multichannel ...Detach cells by incubating with 200 µL TrypLE for 1–2 min. Resuspend cells in 500 µL DMEM complete and transfer... DG8 cartridge into the cartridge holder. Using a 2–50 µL multichannel pipet, load 20 µL of the reaction... -
What is Polymerase Chain Reaction (PCR)
TypeProtocol...PCR tube Ice Bucket 2 μL Template DNA (10 ng-500 ng) 5 μl 10X Taq buffer with MgCl 2 1 μl dNTP mix (10 ... reagents on ice): 2 μL Template DNA (10 ng-500 ng) 5 μl 10X Taq buffer with MgCl 2 1 μl dNTP mix (10 ...normally sterile dH 2 O. To make a 100uM stock of any primer, add a number of µl of dH 2 O equal to the number...annealing temperature step-wise by 1-2°C. The rate of DNA synthesis is ~1-2 kb/min. The extension time can ...work adequately. Divalent cations such as Mg 2+ and Mn 2+ stabilize the buffer solution. These cations...specific instructions about extension time and temperatures. Initial Denaturation for 2 minutes at 94°C. Denature...(PCR). Basic PCR Program Initial Denaturation for 2 minutes at 94°C: This initiation step heats the double... -
Open Resources and Plasmid Tools For Studying C. elegans
TypeBlog Post...unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust GAL4...resources on the Addgene blog Learn more about SapTrap Read about CRISPR/Cas9 in C. elegans Browse our Plasmids...always emphasized the need for open science and collaboration. The field already has comprehensive reference...planned for August 15th, 2019. For more information about the aims and approach of CeNGEN, check out their...elegans. This database provides detailed information about C. elegans 3’UTRs structures, alternative polyadenylation...addition to the wealth of open resources described above, there were many plasmid tools presented at the ...by the worm community. For additional resources about C. elegans and the plasmids we have at Addgene check... -
Science communication: 9 Strategies to Get Your Foot in the Door
TypeBlog Post...more tips on using social media as a scientist here. 2) Public engagement - Participate in science festivals...Sainsbury Laboratory. She is passionate about science communications and loves talking about science, ...Schmidt, a Marie Curie Fellow at The Sainsbury Laboratory. Are you a science student or early career researcher...break into science communication? Everybody goes about this in their own way. The career paths into science...Society gives training in media skills, writing about your research and residential communication. If ... -
Corralling and Cornering the Goldilocks Antibody
TypeBlog Post...non-specific binding in our IHC condition. Fig. 2: Schematic of the goldilocks antibody selection process...Have you ever been stuck googling about an antibody that will be essential to your new research project...high-quality antibodies for IHC. Working with collaborators and support from the Brain Research Through ...extensive library of monoclonal antibodies; Addgene collaborates with these companies to create and provide general...pattern should align with what the atlases (mentioned above) and literature suggest, providing an expectation...open-source database, the mAb3D Atlas will foster collaboration, progress, and innovation by allowing scientists... -
Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015
TypeBlog Post...systems to defend against both DNA and RNA viruses - 2 cas systems regulate at the DNA level and 1 at the... 2015 conference invited me to attend and speak about Addgene’s experience helping scientists share plasmids... with the first keynote presentation. He talked about the impact of the Human Genome project on its 25th...of the meeting was how much we still don’t know about RNA. One talk that really brought this home was ...presented by Todd Lowe from UCSC. Dr. Lowe talked about his work on the 500+ human tRNA genes and, until... Udi Qimron from Tel Aviv University then spoke about using CRISPR/cas to address the growing clinical... -
3D Printing Meets CRISPR Cas9
TypeBlog Post...communicate the nuances of protein structure in a flat 2-D images. In the early days of structural biology,...Until then, you can only wonder what I am talking about. How do researchers use physical models of protein...students work closely with local researchers to learn about their research while creating physical models of...model is an effective teaching tool. You can read about Jacob’s initial reaction to this model on his blog...will wake up in the middle of the night dreaming about the conformational gymnastics that this protein ...Additional Resources on the Addgene Blog Learn All about CRISPR Use CRISPR for Genome Visualization Tips ... -
CRISPR 101: Mammalian Expression Systems and Delivery Methods
TypeBlog Post... already have an end product in mind, steps 1 and 2 will generally be straightforward. Likewise for step...directly back to your specific goal. When thinking about step 3, however, you may be surprised at the number...Delivery methods for mammalian cell lines As mentioned above, the expression system you choose in many ways dictates.... The user should review the current literature about their preferred model. If your expression system... Check out our CRISPR Featured Topic Page Learn about CRISPR Delivery Using Ribonucleoproteins (RNPs) ... -
Adapting Toehold Switches to Detect Zika Virus
TypeBlog Post.... Find Plasmids from the publication at Addgene 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed...allowed gene circuits to be deployed out of the laboratory in a cell-free and biosafe format that was stable...circuit-based sensors to be used outside of the laboratory. The detection threshold for these RNA sequences...sequences (3 nM), however, was quite high and above what would be expected in patient samples (e.g.: blood... -
Tips for Using BLAST to Verify Plasmids
TypeBlog Post... to remove this default setting. Timesaving Tip #2: blastx searches are inherently slower than blastn...such as full plasmid sequences provided by the laboratories that deposit their plasmids with us or other...mismatches and gaps in the alignment. If you are curious about the differences in the blastn programs, check out...Inside Look at NGS Plasmid Quality Control Learn about our Snapgene-powered plasmid maps. Resources on... -
Hot Plasmids - October 2020
TypeBlog Post...products have distinct localizations. Figure 2: Split-wrmScarlet labeling proteins with different... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here. Here's...fluorescent proteins coupled with Fibronectin intrabodies generated with mRNA display (FingRs), Xue Han... -
Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools
TypeBlog Post...C-terminus of human codon-optimized cryptochrome 2 (CRY2) from Arabidopsis thaliana. When exposed to ... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here. ...protein activity while also capturing information about spatial and temporal organization. The design use... -
Split-BioID: An Improved Method for Studying Protein-Protein Interactions
TypeBlog Post...dimerization-dependent protein interactions. FEBS letters, 591 2, 415-424. PubMed PMID: 28032891. This is a different...Addgene Blog Learn about techniques for visualizing protein turnover Learn about plasmids for in vivo... -
CRISPR in the Clinic
TypeBlog Post... Cure for HIV/AIDS. Int J Mol Sci. 2023 Jan 13;24(2):1563. doi: 10.3390/ijms24021563 Resources on Addgene.org...trials The CRISPR therapies don’t stop with the list above! There are many, many more in the earlier stages... and others we just don’t have the time to talk about. Other treatments currently in clinical trials include... -
Personal Protective Equipment (PPE) for BSL-1 and BSL-2 Labs
TypeProtocol... biosafety level 1 (BSL-1) and biosafety level 2 (BSL-2). The BSL-1 classification is for labs working...BSL-1 and BSL-2 labs. Protocols... Protocols Personal Protective Equipment (PPE) for BSL-1 and BSL-2 Labs Personal ...Personal Protective Equipment (PPE) for BSL-1 and BSL-2 Labs Intro to the Lab Bench Check out more protocols ...individuals from potential accidents such as spills. BSL-2 is different because it includes labs that work with...agents associated with diseases in healthy humans. BSL-2 includes all of the precautions needed in BSL-1, however...and/or face shields can be used as needed. For BSL-2 work always wear glasses/goggles in addition to the... -
A Look at Addgene's QC Process
TypeBlog Post...contain the expected plasmid features. Figure 2. Addgene’s basic QC process. Created with BioRender.com...results, and subsequent conclusions. Being vigilant about these errors is an important part of maintaining... -
Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More
TypeBlog Post... Werner S, Marillonnet S. PLoS One. 2011 Feb 18;6(2):e16765. doi: 10.1371/journal.pone.0016765. PubMed...think about how much that’s on it are plants, and the animals you eat ate plants too. Think about how many...is Head of Synthetic Biology at the Sainsbury Laboratory, where she often feels more like an engineer ..., or pathogens to their hosts. I spoke with her about what motivates her research, the MoClo Kit she and...parts for plant synthetic biology Addgene: Tell me about the Golden Gate Modular Cloning (MoClo) Kit. What...make a 10-gene construct, you don’t have to worry about repeating elements anymore. The kit contains sequences...easy to express genes in, which makes it a good laboratory workhorse. Tips & tricks Addgene: Do you have... -
CRISPR 101: Ribonucleoprotein (RNP) Delivery
TypeBlog Post...PMID: 27151215. PubMed Central PMCID: PMC4938170. 2. Zuris, John A., et al. "Cationic lipid-mediated delivery...scientists to make genomic alterations, bringing about a revolution in genome engineering, with new techniques...available by both the Jacob Corn and Alex Schier laboratories. Delivering Cas9-gRNA ribonucleoproteins An...delivery of RNPs is electroporation (A in the figure above), which generates pores in the cell membrane, allowing...as lipid-mediated transfection (B in the figure above), are in the early stages of development. David ...proteins harboring receptor ligands (C in the figure above), which result in the internalization of Cas9-gRNA...show promise for in vivo studies, little is known about the immunogenicity and stability of the Cas9 protein... -
xCas9: Engineering a CRISPR Variant with PAM Flexibility
TypeBlog Post...) 12 (>100) xCas9(3.7)-ABE 69 (1.4) 43 (2) 21 (3) Not examined 16 (>100) Average percent...Blog Learn about Engineered Cas9 PAM Variants Read about CRISPR Base Editing Learn about Homology Directed...editing specificity. We’re excited to learn more about xCas9 - here’s what we know so far! Why do we ...point mutations, also has a strict editing window about 15 nt upstream of the PAM. Even if there’s a PAM...in mammalian cells, finding xCas9 3.7 displayed about the same editing rate as SpCas9 at an NGG PAM. When...are exciting, there’s a lot we still don’t know about xCas9. “I’m not 100% sure xCas9 is going to be flat...Find CRISPR Plasmids for Mammalian Systems Learn about CRISPR History ...