Skip to main content
Addgene

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

Information

In Category

We narrowed to 731 results for: Sis;

Showing: 281 - 320 of 731 results

  1. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    ...as LEDs can be engineered to deliver precise, consistent, and controllable pulses of light. Complicated... heading of two-component systems (TCSs). TCSs consist of a histidine kinase that phosphorylates a response...Leakiness was also an issue; some promoter activity persisted in the inactive state. The dynamic range, defined...

  2. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    ...since become a gold standard in gene expression analysis and a luciferase gene (one of many available to...photon emission is measured resulting from the catalysis of a chemical reaction requiring luciferin, ATP...light-producing reaction. Many luciferase assays require cell lysis as the most efficient means to disrupt the cell ...

  3. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...plasmids articles. These brief articles provide a synopsis of a plasmid or group of plasmids' functions and... used a combination of deliberate and random mutagenesis to isolate the best RFP. They targeted known ...-induced mutations in ATP1A1 can render cells resistant to the small molecule ouabain - and editing at...carefully designed to allow for easy removal of the resistance cassette via Cre recombination. Each knockout...

  4. Antibodies 101: ChIP

    Type
    Blog Post
    ...upsteam sample collection process or the downstream analysis process. And ChIP itself begins at...  Crosslinking...1000 bp. This is to ensure that your downstream analysis focuses on the DNA your protein is actively interacting...complexes. Your DNA is now ready for downstream analysis via any methods suitable for isolated and purified...

  5. Sweating the Small Stuff: Details in the Lab

    Type
    Blog Post
    ...Soon you’ll figure out what prompts you need to consistently and accurately run a protocol.   Figure...hasn't. Developing simple habits like these can consistently reduce errors as you work, and the possibilities...maybe especially) suggestions you are initially resistant to. Just like any other experimental approach,...

  6. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    ...molecular switches in signal transduction pathways and consist of a trimer of three subunits: alpha (Gα), beta...ligand-bound GPCRs of interest. The TRUPATH plasmid kit consists of 20 plasmids encoding 14 alpha, 2 beta, and ..., GPCRs were first mutated via site-directed mutagenesis to bind non natural ligands. These non natural...

  7. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    ...monoclonal Cas9 background because you’ll have more consistent levels of Cas9 expression between experiments...pAAV-hSyn-DIO-hM4D(Gi)-mCherry Our current AAV inventory consists almost entirely of viruses expressing DREADDs ... References 1. Aschauer DF, Kreuz S, Rumpel S. Analysis of transduction efficiency, tropism and axonal...

  8. Plasmids 101: Blue-white Screening

    Type
    Blog Post
    ...way to narrow down candidates for more specific analysis, like PCR or restriction digest. Make sure you...colonies that not only have your plasmid (antibiotic resistance), but also confirm those plasmids have your insert...plasmid-containing cells without using antibiotic resistance. These rely on cell lines susceptible to or dependent...

  9. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    ...Cell and Regenerative Biology is to understand the basis for cardiovascular and metabolic human diseases.... activator-like effector nuclease (TALEN) kit consisting of 834 plasmids. The researchers showed they ... or using a homologous template provided by the sister chromosome. As cells take those two ends and put...

  10. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ...in vivo, they have higher reproducibility and consistency in the final product. Because the constant (non-epitope... as ankyrin repeat proteins. DARPins typically consist of 3-4 repeats, have high specificity and binding...Targeting HER2/neu and CD16 Exhibit Improved Tumor Lysis When Placed in a Divalent Tumor Antigen Binding ...

  11. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ...bumble bee and honey bee microbiome. The toolkit consists of modular parts that are assembled with Golden...plasmids and suitability of various antibiotic resistance markers, promoters, and inducible gene expression...into how the microbiome responds to infection, dysbiosis, and antibiotic treatment. To facilitate these...

  12. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ...this problem, gRNAs can be chemically modified to resist degradation while still effectively serving their...using commercially available kits or ordered as synthesis modifications when purchasing your gRNA. Pro tip... mismatch discrimination and is more nuclease-resistant, resulting in less off-target events. While both...

  13. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    ...your barcoded gRNA library sequences via oligo synthesis using the format indicated below and pool-clone... Wong et al 2015. Alan Wong is currently an Assistant Professor at the School of Biomedical Sciences...Timothy K. Lu. "Enhanced killing of antibiotic-resistant bacteria enabled by massively parallel combinatorial...

  14. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    ...Performance When targeting single genes, SAM consistently shows the highest levels of gene activation ...which are relatively large and are not expressed consistently throughout cells. VPR Description VPR fuses ... dCas9 to activate transcription. This complex consists of the VP64 activator used in other CRISPR activation...

  17. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...categories: probes and editors. A probe array may consist of at least two sensor modules targeting a specific... which can be easily visualized.  The tool-kit consists of highly useful vectors compatible with Gibson...dCas9 expression and contain different antibiotic resistance markers. New CRISPR pooled library from Keith...

  18. CRISPR in the Clinic

    Type
    Blog Post
    ...stem cells isolated from two patients with cystic fibrosis (Schwank et al). They were able to demonstrate...SCD in 2022 as well. Hereditary transthyretin amyloidosis (hATTR) hATTR is caused by a variety of single...Cas9 in Intestinal Stem Cell Organoids of Cystic Fibrosis Patients." Cell Stem Cell. 13, 653-658 (2013)....

  19. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    ..., or other. Bacterial Resistance Enter bacterial antibiotic resistance encoded in the plasmid that...Cloning Method Choose from: Gateway cloning, gene synthesis, Gibson cloning, ligation-independent cloning,...

  20. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    ...graduate students who work as teaching or research assistants to join unions. The topic of unionization continues...of Howell Communications Philip Brenner, PhD - Assistant Professor of Sociology. University of Massachusetts...PhD - Biotechnology Communications Specialist at AsisChem Inc. and Social Media Content Specialist at Thermo...

  21. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...gene networks for the detection of antibiotic resistance and strain-specific diagnostics for the Ebola...Pardee and Alexander Green! Keith Pardee is an Assistant professor in the Leslie Dan Faculty of Pharmacy...the environment.     Alexander A. Green is an Assistant professor at the Biodesign Center for Molecular...

  22. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...silencing of self-genes. Dr. Mello presented a hypothesis that these pathways are involved in RNA based...address the growing clinical problem of antibiotic resistant bacteria. See the picture to the left for his ... vectors are engineered to replace the drug resistantance genes in the bacteria thus making the bacteria...

  23. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ...living cells. Essentially this reporter construct consists of one substrate (the histone-derived peptide)... of these sites was controlled, the reporter consistently showed significantly different FRET levels between...complex or multifactorial diseases, enabling faster diagnosis and treatment and improving patients lives. References...

  24. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ...-transformed cells yielded rates of targeted mutagenesis similar to those achieved by isogenic AdVs encoding...Elsevier, pp 39–67 Ding Q, Strong A, Patel KM, Ng S-L, Gosis BS, Regan SN, Cowan CA, Rader DJ, Musunuru K (2014...CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array of human cells. Sci Rep 4:...

  25. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ...results you need.  Whichever method you choose, resist the temptation to over interpret your results: .... PMID: 15833050. Ma L, Chung WK. Quantitative analysis of copy number variants based on real-time LightCycler...amplification of Plasmodium falciparum multidrug resistance-1 gene (pfmdr1). J Parasitol. 2011 Oct;97(5):...

  26. Plasmids 101: Protein Expression

    Type
    Blog Post
    ...refers to the second step of this process: the synthesis of proteins from mRNA and the addition of post-translational...Overall the general strategy for protein expression consists of transfecting cells with your DNA template of...the infected host cell is eventually lysed. Cell lysis halts protein production, but there are non-lytic...

  27. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...cloning rings to isolate colonies of antibiotic-resistant cells; during the time required to grease, place... below) is simple: After obtaining antibiotic-resistant colonies, use a Sharpie to circle 12-48 individual...specific wells from the 48-well plate for further analysis. To assess CRISPR/Cas9 knockout efficacy, we have...

  28. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...knowledge of protein structure and saturation mutagenesis. mK-GO matures from green to red, and the FT ...absorption spectra). Subach et al’s directed mutagenesis also produced the three separate timers shown...Age-dependent preferential dense-core vesicle exocytosis in neuroendocrine cells revealed by newly developed...

  29. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ...able to study many cellular processes, including mitosis, mitochondrial fission/fusion, nuclear import, ...several groups have designed modular toolkits to assist with tagging genomic loci in other model systems...including mammalian cells. CRISPaint (CRISPR-assisted insertion tagging), developed by the Hornung Lab...

  30. Plasmids 101: Degron Tags

    Type
    Blog Post
    ...one.   SMASh Small molecule-assisted shutoff (SMASh) functions on the basis of both a degron and a viral...Given its popularity and widespread usage, a proteolysis-targeting chimera (PROTAC) was developed for ...

  31. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ...adapted for a wide variety of applications - as mutagenesis tools for genetic studies (e.g. using mariner...Insertion of Transposable Elements by Guide RNA-Assisted Targeting) by the Sternberg group, and CAST (CRISPR-associated...SE, Sternberg SH, Fernández IS (2020) Structural basis of DNA targeting by a transposon-encoded CRISPR–...

  33. Great Results Start with Great Standard Curves

    Type
    Blog Post
    ...to 1:32. Each individual step of the series is consistent at 1:2. When preparing a dilution series, use...non-linear and you should use a different method of analysis. Different fields and assays have varying criteria...run.  When setting up your dilution series, use consistent dilution factors, change pipette tips regularly...

  34. MXS Chaining

    Type
    Blog Post
    ...the flanks of each ligation product, which is the basis for the repeatability of the method. Note that the... 4. Josse J, Kaiser AD, Kornberg A. Enzymatic synthesis of deoxyribonucleic acid. VIII. Frequencies of...Swartz MN, Trautner TA, Kornberg A. Enzymatic synthesis of deoxyribonucleic acid. XI. Further studies ...

  35. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...the detection of rare sequences and single cell analysis. It is also being used in the detection of microbes... al. "Detection of circulating Mycobacterium tuberculosis-specific DNA by droplet digital PCR for vaccine...vaccine evaluation in challenged monkeys and TB diagnosis." Emerging microbes & infections 7.1 (2018): 1...

  36. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...non-dividing cells. The first lentiviral vector system consisted of three plasmids: the packaging, envelope, and...other reporters, and they showed that expression persisted for at least thirty days. Although adeno-associated...integrate into the genome, they could promote oncogenesis by altering local gene expression. Self-inactivating...

  37. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    ...minigenome, the viral polymerase initiates RNA synthesis at the 3’ end of the minigenome to produce the...These products can be detected by Northern blot analysis. In addition, the polymerase will use the minigenome...mRNA can be detected directly by Northern blot analysis or RT-PCR. Uses of the minigenome system Minigenome...

  38. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...tracrRNA, crRNA guides are only ~42 nt long. Direct synthesis of these crRNAs should be significantly cheaper...rates, determined using sophisticated genome-wide analysis. At many of its computationally predicted off-...at Addgene. 3. Kim, Daesik, et al. Genome-wide analysis reveals specificities of Cpf1 endonucleases in...

  39. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    ...immunoprecipitation (locus-specific ChIP). These methods consist of insertional chromatin immunoprecipitation (iChIP...on development of technologies for biochemical analysis of genome functions such as transcription and ...technologies. The locus-specific ChIP technologies consisting of iChIP and enChIP are recent examples. We also...

  40. Hot Plasmids - August 2020

    Type
    Blog Post
    ...CRISPR corner New from the viral service Faster catalysis after yeast-based protein evolution By Joanne ...evolution platform to generate TEV protease variants. Catalysis by TEV protease allows a transcription factor ...CRISPR/Cas Toolkit for plants includes 95 plasmids consisting of CRISPR/Cas nucleases, base editors, gRNA backbones...
Showing: 281 - 320 of 731 results