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Showing: 401 - 440 of 731 results
  1. Lab Automation at Addgene

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    Blog Post
    ...fills all those vials? Well, it used to be a lab assistant, but with increased plasmid demand it was too ...fill 400 vials in 35 minutes compared to a lab assisstant filling about 200 vials in 45 minutes by hand...
  2. Hot Plasmids - October 2020

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    Blog Post
    ...fluorescent timer emits blue light around the time of synthesis and, after a few hours, a conformational change...myeloid leukemia and saw that expression of H2B-FT consistently predicted the cell cycle speed across these ...
  3. A Look at Addgene's QC Process

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    Blog Post
    ...topics that may require additional attention during analysis. These include topics such as: locations in a ...on Addgene.org Educational Resources Sequence Analysis of a Plasmid Protocol ...
  4. Making CRISPR Plasmids Using Fragmid

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    Blog Post
    ...All destination vectors contain an ampicillin resistance cassette and an origin of replication, which ...pRDB_052 (for stable fly cell lines; Hygromycin resistance gene; for genome integration by spontaneous insertion...
  5. 5 Tips for Troubleshooting Viral Transductions

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    Blog Post
    ... will be selectable with the antibiotic whose resistance gene is encoded in the viral vector. This antibiotic... may be some low-level fluorescent protein or resistance marker expression following transfection, higher-level...
  6. Cpf1 Update: Comparison to Cas9 and NgAgo

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    Blog Post
    ...rates, determined using sophisticated genome-wide analysis. At many of its computationally predicted off-... Hur JK, Been KW, Yoon SH, Kim JS. Genome-wide analysis reveals specificities of Cpf1 endonucleases in...
  7. Plasmids 101: Terminators and PolyA signals

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    Blog Post
    ...rho-independent. Rho factor is a helicase which assists RNA polymerase in the termination of the transcript...: PMC102582. Additional Resources A detailed analysis of yeast terminators of varying strength. A discussion...
  8. The Materials Science of Optogenetics Experiments

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    Blog Post
    ...will be erratic, opsin activation/inactivation inconsistent, and behavioral results impossible to interpret...sequentially increasing fineness helps to maintain a consistent stream of light. Test your implant by connecting...
  9. Hot Plasmids and Viral Preps - May 2021

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    Blog Post
    ...compartment was used to optimize D-glucaric acid biosynthesis. This MPyV system encapsulated a rate-limiting... a compact Cas9 ortholog from Staphylococcus lugdunensis that recognizes a NNGG PAM and has comparable...
  10. Tips for a 1st time AAV user (by a Rookie AAV user)

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    Blog Post
    ...the cell pellet, I lyse the cells with this AAV lysis buffer and freeze/thaw the lystate four times in...-on time and 2 hours for the qPCR run and data analysis. Refer to this Addgene AAV Titration by qPCR Protocol...
  11. Degrading DNA with Cascade-Cas3

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    Blog Post
    ... increase efficiency of these systems, though consistent expression can still be a challenge. A multitasking...Dolan, A. E., Liao, M., & Ke, A. (2018). Structure basis for RNA-guided DNA degradation by Cascade and Cas3...
  12. Viral Vectors 101: Voltage Indicators

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    Blog Post
    ...Domain (VSD) GEVIs Simply put, VSD-based sensors consist of a VSD fused to a single fluorescent protein ... Single Fluor Chemigenetic Sensors This system consists of a genetically expressed enzyme, typically on...
  13. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ... easier, and more comprehensive thanks to the consistent development and improvement of CRISPR-based technologies...& Platt, R. J. (2023). Transcriptional linkage analysis with in vivo AAV-Perturb-seq. Nature, 622(7982...
  14. Viral Vectors 101: Producing Your rAAV

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    Blog Post
    ...the virions out. We recommend using a Tris-based lysis buffer and sonication.  The obtained virions from...can then remove the density gradient material by dialysis/buffer exchange and further concentrate the virions...
  15. Antibodies 101: Flow Compensation

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    Blog Post
    ...you can adjust compensation values later in your analysis, unlike voltages.  Figure 1A shows a single-color...always change your compensation values during the analysis portion of your experiment. So, as long as you...
  16. Twenty Years of Addgene Sharing: CRISPR

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    Blog Post
    ...requesting each year (Figure 2). We did a similar analysis with CRISPR pooled libraries, although we used... from this data is that CRISPR technology is consistently showing rapid development and quick adoption...
  17. Google Forums Round Up: First Impressions of NgAgo

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    Blog Post
    ...difficult” GC-rich regions of the genome, which seem resistant to Cas9 cleavage (1). Both the CRISPR-Cas9 and...however rely on cloning into a vector or in vitro synthesis, which could be less desirable if one has to target...
  18. Plasmids 101: Multicistronic Vectors

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    Blog Post
    ... a reporter, such as a fluorescence gene or a resistance gene. These reporters allow you to easily screen...thought to function by making the ribosome skip the synthesis of a peptide bond at the C-terminus of a 2A element...
  19. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ...cytosine deaminase. Three rounds of rational mutagenesis on the ADAR2DD residues that contact the RNA ...RNA-sequencing, the authors performed rational mutagenesis and identified that a S375A mutation in RESCUE...
  20. DNA Purification Without a Kit

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    Blog Post
    ...advantages: they are convenient and provide uniform, consistent results. But they are also less accessible due...columns from DNA purification kits. Kit-free alkaline lysis plasmid miniprep Starting material: 2 mL bacterial...
  21. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

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    Blog Post
    ...Fluorescent Protein? GFP is a ~27 kDa protein consisting of 238 amino acids derived from the crystal jellyfish...began engineering new versions of GFP through mutagenesis in order to improve its physical and biochemical...
  22. AAV Titers: Where Do They Come From And What Do They Mean?

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    Blog Post
    ...image shows that the vast majority of the vectors consist of full particles (white arrowhead) relative to...Droplet Digital PCR versus qPCR for gene expression analysis with low abundant targets: from variable nonsense...
  23. Transferable Skills Guide: Creativity

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    Blog Post
    ... well received, understand that people can be resistant to change. Ask your peers or your superiors to...to explain their rationale when they’re resistant to new ideas. If their answer is “we’ve always done ...
  24. Viral Vectors 101: Preparing Pooled Libraries

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    Blog Post
    ...screening, followed by sequencing and further data analysis. Based on Figure 31.5.2 from https://pubmed.ncbi.nlm.nih.gov...scripts provided by the depositing lab for data analysis.  I'm often asked whether this step can be skipped...
  25. Plasmids 101: Codon usage bias

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    Blog Post
    ...alternative tRNAs Codon optimization With low cost DNA synthesis, one of the primary ways researchers solve the...created codon optimization algorithms and DNA synthesis companies such as IDT and GenScript host online...
  26. Deciphering the Mysteries of Behavior with Viral Vectors

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    Blog Post
    ... (BDNF). BDNF is a small protein involved in neurogenesis, neuronal survival, and synaptic plasticity....Groves, J. O. "Is it time to reassess the BDNF hypothesis of depression?." Molecular psychiatry 12.12 (...
  27. Hot Plasmids - November 2023

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    Blog Post
    ...Susanna Stroik The Liu lab recently used phage-assisted continuous evolution and protein engineering to...  Doman, J.L., Pandey, S., et al. (2023). Phage-assisted evolution and protein engineering yield compact...
Showing: 401 - 440 of 731 results