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Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 328 results for: hal.1

Showing: 161 - 180 of 328 results

  1. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...homologies. Abbreviated protocol for PITCh Step 1: Generate microhomologies in the PITCh vector ~20...editing possibilities MMEJ can enable. References 1. Sakuma, Tatsushi, et al. (2016).  “MMEJ-assisted ...CRISPR-Cas9 with the PITCh systems.” Nat Protoc. 11(1): 118-33. PMID: 26678082. Find the plasmids from ...cell lines can be created in about a month and a half, without the need for complicated cloning of homology...

  2. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...mL polybrene (µL) 0 0 500 1:5 300 200 1:10 150 350 1:50 30 470 1:100 15 485 1:500 3 497 Add 0.5 mL of a... if an MOI >1 was used, some cells may have 1 copy of the transgene, while others have >1 copy of the ... well by pipetting or inverting the tube. Aliquot 1 mL of cell suspension (i.e., 50,000 cells) into each... early polyclonal populations. Sample Data Figure 1: Generation of monoclonal cell lines from expansion...transduced with lentiCas9-Blast and then selected with 1 µg/mL blasticidin for 9 days. Single cells were ...genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature Methods. 2014 Aug;11(8):783-...

  3. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...Bacillus. Appl Microbiol Biotechnol. 2019 Jan;103(1):303-313. doi: 10.1007/s00253-018-9485-4. Epub 2018...and clonal analysis in Drosophila.” Nat Methods 9(1) (2011): 47-55. PubMed PMID: 22205518. PubMed Central...Methods Mol Biol. 2016;1478:53-78. doi: 10.1007/978-1-4939-6371-3_3. PMID: 27730575; PMCID: PMC5270762. ... and lac promoters.” Proc Natl Acad Sci U S A. 80(1) (1983): 21-5. PubMed PMID: 6337371. PubMed Central...solve the problem of broad QF expression-induced lethality in flies. Riabinina et al. also developed chimeric...

  4. How to Prepare for an Industry Interview

    Type
    Blog Post
    ...company culture.  It’s also a good idea to prepare 1-2 unique questions for each interviewer by looking...to take notes after each interview.   Tailor your 1-hour presentation to your audience Start by asking...research project. In total, the interviews lasted from half to all of the day.  This whole process may seem ...carefully. And even if you don’t have technical challenges, your interviewers might.  You’ll probably still...

  5. Hot Plasmids: Summer 2025

    Type
    Blog Post
    ... high brightness and fast maturation time (Figure 1). See the paper for more details on small differences...mGold2t depending on the application. Figure 1: Super-resolution imaging of actin filaments with ...proteins for extended imaging. Nature Communications, 16(1), 3241. https://doi.org/10.1038/s41467-025-58223-5...perform neuronal tracing, anterograde tracing poses challenges, from non-selective anterograde transport to ...

  6. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...centrifugation (lenti) Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher...will help you decide which is right for you. Table 1: Mammalian versus insect cells for viral production...Come From and What Do They Mean? Overcoming the Challenges of Lentivirus Production Tips for Titering Your...

  7. Hot Plasmids - October 2022

    Type
    Blog Post
    ...protein sequence of the predecessor GEVIs (called ASAP 1 and 2), they found an indicator that is optimized ...several other constructs in plasmid form. Fig. 1: Summary of the approach and outcomes of the screen.... The resultant JEDI-2P sensors overcome many challenges in 2-photon in-vivo imaging of voltage fluctuations...

  8. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

    Type
    Blog Post
    ...Rodríguez et al., 2011).  Genetics of SPARC Figure 1: SPARC and SPARC2 use the GAL4-UAS system along with... several common genetic tools in Drosophila (Fig. 1):  GAL4-UAS, a transcription factor binding site....titration of the AAV. So far, tools to overcome this challenge in Drosophila require heat-shocking the system...

  9. AAV Production in HEK293 Cells

    Type
    Protocol
    ...7.5% Sodium Bicarbonate, 7.5 mL 1 M HEPES to 750 mL DMEM + 1 g/L glucose. 1 mg/mL polyethylenimine (PEI) ...Calculate the amount of each plasmid needed to have a 1:1:1 molar ratio with 2 mg total DNA per CS5 Plasmid ... determine the total μg/bp we need to achieve a 1:1:1 molar ratio of each plasmid: 2000 μg / 24,961 bp...deionized water + 100 mL of 1 M Tris HCl pH 8.5 + 60 mL of 5 M Sodium Chloride + 4 mL of 1 M Magnesium Chloride...high glucose, Corning 10-013-CV DMEM, low glucose (1 g/L) glucose, sodium pyruvate, Corning 10-014-CV (... sodium bicarbonate, Corning 25-035-CI (optional) 1 M HEPES, HyClone SH30237.01 (optional) L-alanyl-L-glutamine...cations can affect the attachment of adherent cells) 1 mg/mL Polyethylenimine (PEI) 25 kDa MW Pro-Tip Other...

  10. Tips for arabidopsis transformation

    Type
    Blog Post
    ...Arabidopsis thaliana seedlings following floral dip transformation’, Plant Methods. BioMed Central, 2(1), p. ...with a single insertion because it will segregate 3:1 in the T2 generation. You also have no control over... transgenic plants in the process.     References 1. Emami, S., Yee, M. and Dinneny, J. R. (2013) ‘A robust...

  11. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ...tethered to bring them into close proximity (Figure 1). Formation of the complete FP results in a fluorescent...detected by fluorescence microscopy. Figure 1: How BiFC works. A split FP (e.g., yellow fluorescent... protein interactions in living cells. Nat Protoc 1:1278–1286. https://doi.org/10.1038/nprot.2006.201 ...also theoretically possible, but likely more challenging, with lentiviral expression. For more detail ...

  12. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...environments such as live cells and tissues (Figure 1). These flexible strategies give you the best of both...majority of in-cell labeling strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion...Follow him on twitter @rhodamine110.   References 1. Liu, Zhe, Luke D. Lavis, and Eric Betzig. "Imaging... include: Self-labeling tags (e.g., SNAP-tag, HaloTag, TMP-tag) – These widely used systems consist of...

  13. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ... fusions that retain their individual activities (1). Researchers also discovered the modular format of...systems could be combined effectively (3). In Figure 1 for instance, the DBD of the fusion protein shown ...ubiquitin system has been devised (8). References 1. Casadaban M.J., Martinez-Arias A., Shapira S.K., ...functional luciferase only when fusions of the two halves of the luciferase protein interact and enable intein...

  14. Summer SciComm: Science Storytelling

    Type
    Blog Post
    .... In literary terms, conflict can be defined as (1) the struggle between two opposing forces or (2) the...journey even when they know the ending.    Fig. 1: We all know that the superheroes win in the end. ... and what can be left out, which can be a big challenge!  The resolution Throughout your conflict, you...

  15. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    ...and take a look at each of its parts. Figure 1: Parts of an AAV transfer plasmid. Parts shown in ..., ubiquitous expression, while the human synapsin 1 promoter is neuronal specific (Haery et al., 2019)...sequencing conditions which can read through this challenging sequence. Transgene The transgene is the gene...

  16. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    ...sequences were initially discovered in E. coli in 1987 (1), the concept that these clustered repeat sequences...neutralize CRISPR/Cas. Earlier this year, expression of 1 of 5 anti-CRISPR genes were found to inactivate the...defense against invading phages. Rising to the challenge - Evolution of anti-CRISPR genes in phages The...

  17. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    ...protein. Did you know that most cephalopods die after they lay their 1 clutch of eggs? Propagation is ...organism blog posts: Five Popular Model Organisms, Part 1 Five Popular Model Organisms, Part 2 ...share all of these new organisms continues to be challenging. Organism specific viral vector systems would...thus making progress on the NSF “G to P grand challenge.” Creating a productive, new scientific community...proteins, and DNA. Euprymna scolopes (Joshua Rosenthal) The Hawaiian bobtail squid has an extreme number...

  18. Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis

    Type
    Blog Post
    ...restriction digests are comprised of 2 separate steps: 1) incubating your DNA with restriction enzymes which...The resulting gel image includes a 1kb ladder (lane 1) that has bands ranging from about 500bp to 10kb, ...yourself or obtained it from a colleague down the hall, it is always a good idea to take some time to confirm...

  19. CRISPR Library Amplification

    Type
    Protocol
    ...Vented Falcon Tubes at 30-37 ℃, 225 rpm for 1 hour. After the 1 hour shaking period, pool and gently mix ...Last Update: August 17, 2023 Workflow Timeline Day 1: Transform, recover, set up overnight growth (Estimated... (Default: 4 tubes of Endura Duos, Lucigen, 60242-1) Alternatives include Stbl4 cells or other ultra-high...0.1 cm ) 20 mL SOC recovery media (Lucigen, 80026-1) 8X LB Agar + Antibiotic 245 mm bioassay plates (Molecular...into each of four 14 mL Vented Falcon Tubes and have 1 mL SOC per electroporation readily available for post-electroporation...autoclaved, sterile reagents for all steps. Procedure Day 1 Add 200 ng DNA to each 50 µL aliquot of thawed Endura...: Bio-Rad Micropulser Ec1 0.1 cm cuvette, 1.8 kV, 1 pulse. Aliquot 25 µL DNA-Endura into pre-chilled cuvette...

  20. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    ...mediate A•T-to-G•C base changes (Figure 1), but it can be challenging to make these base changes, especially...dCas to the genomic DNA of interest  Figure 1. When ABEs are in action they bind DNA so that the...𝛽-globin), which cause sickle cell disease and 𝛽-thalassemia, can be mitigated with the expression of fetal...
Showing: 161 - 180 of 328 results