We narrowed to 836 results for: Eras
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TypeBlog PostPublishedAug. 1, 2023, 1:15 p.m....didn’t need to be strictly followed in order to generate high-fidelity reactions. (Need a refresher? The...cycling conditions (see Table 1). Enzyme Temperature Incubation time/cycling Overhang length ...some help! References Potapov, V., Ong, J. L., Kucera, R. B., Langhorst, B. W., Bilotti, K., Pryor, J...10.1021/acssynbio.1c00525 Pryor, J. M., Potapov, V., Kucera, R. B., Bilotti, K., Cantor, E. J., & Lohman, G...
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Quickest Way to Deposit Plasmids: The Deposit Spreadsheet
TypeBlog PostPublishedMarch 8, 2017, 3:15 p.m....expression, lentiviral, retroviral, AAV, RNAi, luciferase, cre/lox, yeast expression, worm expression, ...Growth Temp Choose from 30 °C, 37 °C, or room temperature. Growth Strain Please indicate which growth...GenBank files. We encourage submission of QUEEN-generated GenBank files (Mori and Yachie, 2021) but can ...we’ll use the information in the spreadsheet to generate plasmid pages. We’ll send you a deposit confirmation... -
Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene
TypeBlog PostPublishedApril 6, 2021, 12:30 p.m.... its analysis can be found here: A Novel Next-Generation Sequencing and Analysis Platform to Assess the...the pool - ex. the gRNA in a CRISPR library. In general, our QC scientists work with the depositing lab...Mueller I, Mitsopoulos A, Fan M (2020) A Novel Next-Generation Sequencing and Analysis Platform to Assess the...Preparations from Viral DNA Extracts. Human Gene Therapy 31:664–678 . https://doi.org/10.1089/hum.2019.277... -
Hot Plasmids and Viral Preps - January 2021
TypeBlog PostPublishedJan. 5, 2021, 2:15 p.m....Synthetic Biology publication, Mary Dunlop’s lab generated a photosensitive Cre, Opto-Cre-Vvd, by splitting...Casɸ-2, and Casɸ-3. This system is notable for several reasons. Casɸ is only ~70kDa, with a molecular ...cultures. HeLa cells expressed a fusion of keratin with mGold (top) and mVenus (bottom). Cells were... variants. It's also one of the smallest AAV-deliverable ABE. Find the plasmid here Jesse Zalatan’s... -
Corralling and Cornering the Goldilocks Antibody
TypeBlog PostPublishedAug. 9, 2022, 1:15 p.m....collaborates with these companies to create and provide general availability of their recombinant antibodies. Since...that the primary antibody is very weak or with general non-specific binding. If the sample differs from...align with what the atlases (mentioned above) and literature suggest, providing an expectation and comparison...imaged pattern does NOT align with the atlases/literature and is unrelated to the target protein, we categorize... -
6 Tips for Grant Writing
TypeBlog PostPublishedOct. 18, 2016, 2:30 p.m....back to you, and for you to make the relevant alterations before submitting - handing them your application...during your career; you learn new techniques and interact with new people. If you are looking to move to...non-scientists. Most fellowships are funded by the general public (either through taxes or charitable donations...an IDP Get Tips on Keeping Up With Scientific Literature ... -
5 Tips for Troubleshooting Viral Transductions
TypeBlog PostPublishedAug. 11, 2016, 2:23 p.m....more than once. Fibronectin is another membrane-interacting protein that can be used to enhance transduction...aren’t working, you can check if virus is being generated by your transfected packaging cells. This check...and transduction. Once you’ve harvested, it’s generally a good idea to titer your virus. Getting an accurate...of primary human T lymphocytes for adoptive immunotherapy." Acute Leukemias VIII. Springer Berlin Heidelberg... -
Illuminating Epigenetics with A FRET Based Biosensor
TypeBlog PostPublishedNov. 19, 2015, 3:30 p.m....involve any changes to DNA sequence (Epigenetics literally means ‘on top of the genome’). Epigenetic modifications...prevents the cellular machinery from physically interacting with the genes in the compacted region and results...can influence the transcription of genes by RNA polymerase II, or by the addition of methyl- groups to the...restriction enzyme digestion, DNA adenine methyltransferase identification, and bisulfite sequencing. ... -
PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs
TypeBlog PostPublishedJune 25, 2024, 1:15 p.m....efficiencies with pridict.it. This approach leverages the strengths of PRIDICT/PRIDICT2.0 while accounting...into play. Nicolas Mathis received his PhD from Gerald Schwank's lab at the University of Zurich (Switzerland...DeepSpCas9, a deep learning–based model with high generalization performance. Science Advances, 5(11), eaax9249..., A., Kuzmin, I., Raudvere, U., Peterson, H., Liberante, F. G., & Parts, L. (2023). Prediction of prime... -
Antibodies 101: Conventional vs Spectral Flow Cytometry
TypeBlog PostPublishedAug. 3, 2023, 1:15 p.m....the beads or cells. Overall, spectral flow cytometry has the ability to generate incredible amounts of...which spectral flow cytometry requires special considerations. Conventional flow cytometry During conventional...student at Mayo Clinic studying host-microbe interactions in early life. Resources More resources ... -
3D Printing Meets CRISPR Cas9
TypeBlog PostPublishedOct. 17, 2017, 12:55 p.m....that allowed researchers to create stunning, interactive images of their proteins. Since the 1980’s, journal...obscures the flexible nature of all proteins in general, and the amazing conformational flexibility that...strand. In addition, this model also displays the several Lys/Ala mutations reported by Ian Slaymaker and...classroom or trying to explain your science to the general public, visit the Center for BioMolecular Modeling... -
Quick Guide to Working with Drosophila Part 1: Getting Started with Flies
TypeBlog PostPublishedJuly 13, 2017, 2:30 p.m.... 25°C. At low temperatures like 18°C, flies take longer to develop. At high temperatures like 30°C, flies...and Y chromosomes are miniscule, so researchers generally do not manipulate them. Essentially, the X, second...the mutation won’t drift away with each passing generation. Another feature of balancer chromosomes is ... -
Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity
TypeBlog PostPublishedOct. 11, 2018, 12:30 p.m....bloggers Becky Kucera, M.Sc. and Eric Cantor, Ph.D. from New England Biolabs. Becky Kucera is a Principal...This post was contributed by guest bloggers Becky Kucera, M.Sc. and Eric Cantor, Ph.D. from New England ... more than 10 modules often rely on two-step hierarchical approaches using different Type IIS restriction... -
Anatomy of a Plasmid Page at Addgene
TypeBlog PostPublishedFeb. 4, 2016, 3:30 p.m....trail at the top of any plasmid page gives the hierarchy of web pages the plasmid page is nested in. You...it? Your best bet is the Plasmid Map: We auto-generate a plasmid map based off of the plasmid sequence...plasmids are sent as bacterial stabs at room temperature. Core of the plasmid page - Core of the plasmid...), antibiotic resistance, and optimal growth temperature are listed, as well as the name of the strain... -
Antibodies 101: Producing Recombinant Antibodies
TypeBlog PostPublishedAug. 13, 2024, 1:15 p.m....polyclonal antibodies are readily available from several sources, fewer sources of recombinant antibodies...appropriate buffer, it’s time to quantify it. There are several options to choose from depending on the time, reagents...2018). Aggregation can arise from a shift in temperature, pH, or salt concentration (Amin et. al., 2014... shipped to customers on ice, preventing any temperature shifts. Of course, we are also cautious during... -
Plasmids 101: Colony PCR
TypeBlog PostPublishedMay 12, 2016, 2:30 p.m....set up a standard PCR reaction (primers, dNTPs, polymerase) using the supernatant of lysed bacteria as template...standard PCR reaction: combine template, primers, polymerase, and dNTPs and then incubate with a standard ...heating step of the PCR reaction. A standard Taq polymerase is sufficient. Controls: Controls can make ...them onto the gel. The figure above summarizes generalized expected results for the three primers previously... -
Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research
TypeBlog PostPublishedJuly 29, 2025, 1:15 p.m....RuvC-like), and Cas12a has one (RuvC-like), TnpB also operates with a single RuvC-like domain. In simple terms..., viral vectors have been successfully used to generate gene-edited plants without the need for tissue... additional enzymes or domains (increasing the overall size), using TnpB as the core nuclease could help...Conclusion & Future Directions TnpB represents a new generation of compact, efficient, and versatile genome editing... -
Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)
TypeBlog PostPublishedJuly 8, 2025, 1:15 p.m....Figure 1 for a visual overview! Figure 1: General overview of the MoClo assembly process to produce... common MoClo nomenclature. The overhangs are generated using type IIS restriction enzymes (BsaI, BbsI...MoClo plate, always do so in cold conditions and preferably on dry ice. Careful use of your glycerol stocks...plates. However, even though the MoClo system is generally very efficient and precise, positive colonies ... -
Plasmids 101: Plasmid Incompatibility
TypeBlog PostPublishedMarch 31, 2020, 1:15 p.m.... are several different versions of the plasmid partitioning system in the bacteria, but generally it consists...coresident plasmid. Asymmetric plasmid loss occurs for several reasons, including one plasmid blocking replication...compatible or not. Using plasmid incompatibility for therapeutic advantage Although it may seem like plasmid incompatibility... -
Overcoming the AAV Size Limitation for CRISPR Delivery
TypeBlog PostPublishedSept. 16, 2020, 1 p.m....multiple gRNAs into separate AAV vectors, increasing overall packaging capacity but necessitating purification...labeling, enrichment on streptavidin and next-generation sequencing). Using this sensitive method, they...that the field has overcome with split AAVs. In general split AAVs break a large transgene in two pieces... Wang D, Zhang F, Gao G (2020) CRISPR-Based Therapeutic Genome Editing: Strategies and In Vivo Delivery...