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We narrowed to 731 results for: Sis;

Showing: 241 - 280 of 731 results

  1. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

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    Blog Post
    ...bacterial resistance marker used to grow plasmid containing bacterial cultures Bacterial Resistance: Use this...terms with gene and guide RNA / shRNA inserts Resistance Marker: This indicates the selectable marker ... this to choose the antibiotic resistance gene(s) used for selection in bacteria. Has Viral Service?: ...

  2. Tips from the Repository Trenches: Using Barcodes to Track Samples

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    Blog Post
    ...across oceans and continents. Each deposit may consist of anywhere from one to hundreds of samples, with...plasmids contain either an ampicillin or kanamycin resistance gene. Every step along the way involves good ...your lab ensure that sample ID’s are unique, consistently labeled, and that numbers are not switched around... with plasmid transformation, preparation, and analysis from our protocols and molecular biology reference...

  3. Plasmids 101: Protein tags

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    Blog Post
    ...purification with glutathione, can also protect against proteolysis. That’s one form of instability. Prokaryotes ...purification with glutathione; protects against proteolysis, but may reduce solubililty HA YPYDVPDYA or...EINAPTDGKV EKVLVKERDA VQGGQGLIKI GVHHHHHH  9 Combo Consists of a bacterially derived in-vivo biotinylation...KEFLDANLAG SGSGHMHHHH HHSSGLVPRG  12 Solubililty Assists in proper folding V5 GKPIPNPLLGLDST   1.4 Affinity...

  4. Your Lentiviral Plasmid FAQs Answered

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    Blog Post
    ...1]; and 2) the potential for oncogenesis through insertional mutagenesis. These risks are largely based...differ in two ways. Firstly, 2nd generation systems consist of an envelope plasmid and a packaging plasmid ...the deletion reduces the risk of insertional mutagenesis by limiting the interaction between viral LTR...

  5. Viral Production at Addgene

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    Blog Post
    ...preparations are subjected to polyacrylamide gel electrophoresis (PAGE) followed by silver staining or SYPRO...purified and non-purified AAV subjected to gel electrophoresis. Viral capsid proteins VP1, VP2, and VP3 are...preparation shows that the vast majority of the vectors consist of full particles (white arrowhead) relative to...transfection. PCR products are analyzed by gel electrophoresis to ensure the correct banding pattern and sizes...

  6. Getting the Most from Your Lentiviral Transduction

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    Blog Post
    ...other cell lines, or infected personnel and can persist undetected; unlike infections with larger microbes...al. found that lentivirus produced at pH 6 are resistant to multiple freeze thaw cycles. Consequently, ... to ensure the best possible transduction. Be consistent Once your lab establishes the best conditions...for your cell lines and virus properly and be consistent when following protocols. Need Virus? Check out...

  7. Important Considerations in Optogenetics Behavioral Experiments

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    Blog Post
    ...rudimentary, but before you do any type of behavior analysis, simply take a few animals and stimulate them ...optogenetics experiment is contained within the hypothesis you are testing. In other words, your optogenetic...explained primarily in two different ways 1) your hypothesis is wrong or 2) there is some problem with the...optogenetic tools derived from direct comparative analysis of microbial opsins." Nature methods 9.2 (2012...

  8. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

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    Blog Post
    ...contributed by guest blogger Joachim Goedart, an assistant professor at the Section of Molecular Cytology...coded by the multiple cloning site and therefore consist of random amino acid sequences or (ii) designed... an inert, unstructured peptide and therefore consisted of glycines, serines and other small, non-aliphatic...blogger, Joachim Goedhart! Joachim Goedhart is an assistant professor at the Section of Molecular Cytology...

  9. Hot Plasmids: Summer 2024

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    Blog Post
    ...loss. Ci Ji Lim’s lab found that adding Late Embryogenesis Abundant (LEA) proteins, small proteins that... by helical LEAs. C-E) cryo-EM single-particle analysis of PRC2 with RvLEAMshort (1:6 molar ratio) with...s other recent deposit builds on Prime-editing-Assisted Site-Specific Integrase Gene Editing (PASSIGE)...CRISPRoff system — a programmable epigenetic editor consisting of dCas9 fused to a DNA methyltransferase — that...

  10. CRISPR 101: Validating Your Genome Edit

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    Blog Post
    ...of WT and edited cell lines. Then decomposition analysis is performed yielding a graph of edits in the ... Fig. 3: Restriction enzyme screening for resistivity to digestion, indicating targeted edits. ...solely for restriction enzyme sensitivity or resistivity, and thus be a marker for edited cells. As with...Software such as CRISPResso can help with the data analysis. If NGS is not an option, you can still detect...

  11. Plasmids 101: Biotinylation

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    Blog Post
    ... are essential for processes such as gluconeogenesis, lipogenesis, amino acid metabolism and energy transduction...Kresege et al., 2004). Avidin is a glycoprotein that consists of four identical subunits that can each bind ...non-selectively label DNA and RNA. Photobiotin acetate consists of a biotin molecule attached to a photoreactive...

  12. Treating Muscular Dystrophy with CRISPR Gene Editing

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    Blog Post
    ...off-target editing. In order for the therapy to persist over time, stem cell editing would be highly desirable...improved, with vastly reduced inflammation and necrosis compared to unedited animals. In grip strength...approach, increasing the possibility that edits will persist long-term. In accordance with this result, Nelson...CRISPR Could Be Used to Treat Cataracts and Cystic Fibrosis Read How Homology Directed Repair is Used to Edit...

  13. How to Design Your gRNA for CRISPR Genome Editing

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    Blog Post
    ..., it would be unwise to make conclusions on the basis of the activity of a single gRNA, and thus diversity...on careful experimental design, execution, and analysis. Many thanks to our guest blogger John Doench....2017.97 Hanna RE, Doench JG (2020) Design and analysis of CRISPR–Cas experiments. Nat Biotechnol 38:813...https://doi.org/10.1038/s41576-018-0059-1 Veres A, Gosis BS, Ding Q, Collins R, Ragavendran A, Brand H, Erdin...

  14. Plasmids 101: Repressible Promoters

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    Blog Post
    ...systems. Binary systems are genetic tools that consist of two parts, both of which must be present for...allows them to be used together for complex genetic analysis in Drosophila or for precise control over multiple...Desplan. “Power tools for gene expression and clonal analysis in Drosophila.” Nat Methods 9(1) (2011): 47-55...transgene expression, lineage tracing, and mosaic analysis.” Cell 141(3) (2010): 536-48. PubMed PMID: 20434990...

  15. Quick Guide to All Things Lentivirus

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    Blog Post
    ...the Moloney Murine leukemia virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing...generation. The 1st generation of lentiviral vectors consisted of three plasmids encoding 1) the lentiviral vector...Efficient gene transfer Potential for insertional mutagenesis: Replication-incompetent lentiviruses with human... from one batch to another. For experimental consistency it’s a good idea to purify and titer your virus...

  16. Transferable Skills Guide: Teamwork

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    Blog Post
    ...of a senior thesis), I knew early on that I wanted to work with a partner. WPI’s emphasis on teamwork ...decision about something? Fix a problem? Test a hypothesis? Whether you officially call it a “project team... risky shots at the goal rather than going for assists that are more likely to lead to scores. That player...

  17. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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    Blog Post
    ...post was contributed by Jordan Ward who is an assistant professor at the University of California, Santa...elegans relied on selective markers such as drug resistance (Chen et al., 2013), fluorescence (GFP; Tzur ...Thank you to our guest blogger! Jordan Ward is an assistant professor at the University of California, Santa...on gene regulation in nematode molting and spermatogenesis. His group also builds tools to improve genome...

  18. CRISPR 101: Cytosine Transversion Editors

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    Blog Post
    .... Like other ABEs, the editor they were using consisted of Cas9 fused to an engineered adenosine deaminase...tool for predicting CGBE editing outcomes and assisting sgRNA selection is available at http://www.sunlab.fun...A., Yang, D., Mok, B., Replogle, J. M., Xu, A., Sisley, T. A., Weissman, J. S., Adamson, B., & Liu, D....developed using CRISPRi screens, target-library analysis, and machine learning. Nature Biotechnology, 39...

  19. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

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    Blog Post
    ...a prestigious institution: This criterion is consistently under scrutiny. USCIS wants you to prove that... is very important in this case.  You can still assist your PI in the peer review process, but remember...love of science and technology, Rachel is proud to assist companies and foreign nationals working in the ...dedicated to the practice of US immigration law and assists individuals, researchers, entrepreneurs, companies...

  20. Editor's Choice, August 2016

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    Blog Post
    ...optimize and troubleshoot the system. Site Directed Mutagenesis by PCR & Single Base Editing with CRISPR For ...technology may make some uses of Site Directed Mutagenesis by PCR obsolete! Honorable mentions Of course... in chronological order here: Site Directed Mutagenesis by PCR Google Forums Round Up: First Impressions...

  21. Who Gives a Tweet? 9 Facts About Scientists on Twitter

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    Blog Post
    ...this article: “A systematic identification and analysis of scientists on Twitter.” This paper addresses..., or are friends, are included in this cluster analysis. The closer two dots are to each other, the more... C. R. (2017). A systematic identification and analysis of scientists on Twitter. Plos One,12(4). doi:...

  22. CRISPR 101: Cytosine and Adenine Base Editors

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    Blog Post
    ... than point modifications. CRISPR base editing persists through multiple cell divisions, indicating that...specificity. In this first, the Liu lab used phage-assisted evolution selection systems to generate ABE8e(... a similar approach. Dali Li’s lab’s A&C-BEmax consists of a fusion of both cytidine and adenosine deaminases... E., Doudna, J. A., & Liu, D. R. (2020). Phage-assisted evolution of an adenine base editor with improved...

  23. Sensing Neuronal Dopamine

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    Blog Post
    ...dopamine. Methods for measuring Dopamine In vivo microdialysis The gold standard method of measuring DA and...neurotransmitters in the brain has been in vivo microdialysis of extracellular fluid (Darvesh et al., 2011...References Darvesh, Altaf S., et al. "In vivo brain microdialysis: advances in neuropsychopharmacology and drug...

  24. FastCloning: A Newer, Simpler Cloning Technique

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    Blog Post
    ... these techniques and is a multi-step process consisting of digestion, purification, ligation, and transformation... 1. Celie, Patrick HN, Annabel HA Parret, and Anastassis Perrakis. "Recombinant cloning strategies for...Plasmid Cloning by PCR Perform Site Directed Mutagenesis by PCR Catch up on Your Plasmid Background with...

  25. Sleeping Beauty Awakens for Genome Engineering

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    Blog Post
    ...social and regulatory reasons. 1.) The transposon consists of a mirrored set of inverted repeats (green) ...1) an SB transposase, the enzyme required for catalysis of transposition; and 2) a transposon containing...determined by transposition assays of antibiotic-resistance genes conducted in HeLa cells). In their Nature...

  26. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

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    Blog Post
    ...culture, only the plasmid with the antibiotic resistance gene and origin will replicate, and the transgene...recombined vector backbone contains the antibiotic resistance gene (AmpR) and origins of replication (ori and...of bacterial plasmid DNA: electron microscopic analysis of in vitro intramolecular recombination. Proceedings...

  27. Single-cell tracking of lineage and identity with CellTag

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    Blog Post
    ...labels cells with heritable barcodes. Barcodes consist of an 8bp variable region and a 6 bp fixed region... a sample of cells was collected for scRNA-seq analysis and the remaining cells were maintained in culture...detailed CellTag protocol on protocols.io, code and analysis tutorials on GitHub, and a CellTag lineage data...

  28. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

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    Blog Post
    ...was then screened through successive rounds of mutagenesis for the brightest teal fluorescent proteins that...scientists from Florida State University, they used a mutagenesis approach and found that specific amino acid changes...protein localized to their expected locations. An analysis of mWasabi gave similar results. We hope this ...

  29. CRISPR Challenges: Standardization and Homology Directed Repair

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    Blog Post
    ...much more about cellular DNA repair pathways. An emphasis on cellular repair post-CRISPR cleavage When I...Ribonucleoprotein (RNP) Delivery The RNP delivery method consistently leads to high on-target editing and low off-...strategies activate different HDR sub-pathways like synthesis-dependent strand annealing (SDSA) and single-strand...

  30. In Vivo Biotinylation of Bacterial Fusion Proteins

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    Blog Post
    ...GST-glutathione. Biotin-based immobilization for kinetic analysis Proteolytic removal of the fusion protein could...could provide a more relevant kinetic analysis, but the trick is to utilize a second immobilization method... purification strategy outlined here provides a basis to determine binding kinetics using a multitude ...

  31. Science Careers: Unruly Interests Feed Many Paths

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    Blog Post
    ...replication and gene transcription during vertebrate embryogenesis. My interests, though, have proven unruly. Instead... at Science magazine. That one I just could not resist applying for. Not that I knew anything about being...like word choice, punctuation, and stylistic consistency. If you’ve ever doubted the importance of copy...

  32. Plasmids 101: TOPO Cloning

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    ... Blog Perform Site Directed Mutagenesis by PCR Use REPLACR Mutatgenesis to Easily Generate Insertion ...approach to molecular cloning and polynucleotide synthesis using vaccinia DNA topoisomerase. Shuman S. J ...

  33. Plasmid Cloning by PCR

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    Blog Post
    ...molecular cloning consist of: Leader Sequence: Extra base pairs on the 5' end of the primer assist with restriction...selecting restriction sites, you should use a DNA analysis tool, such as Addgene’s Sequence Analyzer, to ...

  34. Improving Transduction Efficiency with Exosome AAVs

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    Blog Post
    ... often describe a process that begins with cell lysis to release AAV particles with subsequent purification...glioblastoma). They also found that exo-AAVs were more resistant to neutralizing anti-AAV antibodies. This is particularly...areas that are difficult to reach, and are more resistant to inactivation through anti-AAV antibodies. ...

  35. Creating accessible biology activities in schools with BioBits

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    Blog Post
    ... way into high school classrooms, like gel electrophoresis to separate DNA or transforming cells with ...practice with practical lab skills like developing a hypothesis, precisions pipetting, and good record keeping...in functional genomics research Learn how the Arabidopsis research center brings plant science to life ...

  36. "Build Your Plasmid": The Game - Play to Teach

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    Blog Post
    ...about Synthetic Biology Find Arabidopsis educational resources at the Arabidopsis Biological Resource Center...Promoters, Inserts, Tags, Origins of Replication, Resistance gene and Selection Markers) and one “Transfer...

  37. Viral Vectors 101: Inverted Terminal Repeats

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    Blog Post
    ...required for replication, packaging, and vector persistence. They work in conjunction with the viral rep ... they flank the gene of interest. A single ITR consists of palindromic arms (A-A’, B-B’, and C-C’) that... also helpful in quantifying ITR mutants since analysis of individual reads allows for identification ...

  38. Multiplexed Overexpression of Regulatory Factors (MORF) collection

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    Blog Post
    ...the human fetal expression atlas. Computational analysis was used to infer differentiation trajectories... the 24 TFs they tested, 22 expressed markers consistent with the cell type that was predicted based on... if the differentiated cells had morphologies consistent with the predicted cell type (e.g., axon-like...

  39. Cultivating Community Science at BosLab

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    Blog Post
    ...space where BosLab exists today. Back then, it consisted of “wooden slab benches” set on top of filing ...companies and BosLab has grown into an organization consisting of over twenty members and volunteers. Transforming...BosLab. Once he optimizes plant cell culture and transgenesis he wants to share his knowledge with the community...

  40. Plasmids 101: NGS Quality Control for Pooled Libraries

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    Blog Post
    ...of other analysis scripts available, some associated with specific libraries, but any analysis script should...hundreds of thousands if not millions of reads, this analysis must be done in an automated manner. Addgene commonly...
Showing: 241 - 280 of 731 results