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 210 results for: cycs

Showing: 1 - 30 of 210 results

  1. Bringing Sustainable Practices to the Lab: Recycling

    Type
    Blog Post
    ... or city recycling services don’t offer plastic recycling, specialized companies can recycle plastic--...and to print a free shipping label. Recycle nitrile gloves Terracycle is a company that takes items not ... blog posts. This first post will be based on recycling! With the popularity of single-use items, plastic... generation. Less than 9% of plastics are ever recycled, so adopting a few meaningful changes in the lab...environmentally friendly. Adopt one or all of the recycling practices below. Reuse lab cooler boxes Some vendors...sticker off the box, remove the ice pack (it’s not recyclable), tape the box shut, and drop off the box at ... United States Postal Service picks up mail. Recycle plastic packaging Packaging is a huge contributor...

  2. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ... particle — and of the normal, replicative life cycle — can help you understand these seemingly odd choices...vectors. Key to the wild-type virus’ reproductive cycle is the fact that the virus must have its structural...as the viral particle emerges. Retrovirus life cycle Retroviruses go against the grain of the Central...will discuss several aspects of the viral “life cycle” in order to understand what these vector particles... cells (Fig. 3).     Figure 3: The life cycle of a retrovirus. Image courtesy of Jeremy Luban,...Luban, MD. Figure 3 depicts the viral life cycle. One should ponder this figure often, especially the ...particles. And there you have it, the full “life cycle” of expression of selfish, lazy genes of a non-living...

  3. Tetracycline Inducible Expression

    Type
    Collection
    ...e.g. tTA). Use of tetracycline or a derivative A derivative of tetracycline, doxycycline (dox), is a preferred...Collections Tetracycline (Tet) Inducible Expression Tetracycline (Tet) Inducible Expression Additional Resources...Collections Background Tetracycline Off Tetracycline On Experimental Tips Tet Plasmids Background To advance the...induction with efficient tetracycline uptake. Search our collection for all tetracycline plasmids. Tet System...19 nucleotide tetracycline operator (tetO) sequence, and is recognized by the tetracycline repressor (tetR...endogenous bacterial system, if tetracycline, or one of its analogs like doxycycline, are present, tetR will ...will bind to tetracycline and not to the TRE, permitting transcription. Tetracycline-dependent promoters ...

  4. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...uracil-excision based (USER) sites, flanked by ADH1 and CYC terminators for easy cloning of genes and promoters...unique tissue-specific promoters expressing GFP, tetracycline response elements, and shRNAs many of which ...control of the TRE promoter, which allows for doxycycline-inducible expression. The end result is genomic...controlled expression of the dCas9-VPR transgene by doxycycline in your target cells. This plasmid adds to the...gondii infection, and necessary for the asexual cycle of malaria parasites. Find Lourido CRISPR Plasmids...

  5. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    ...C cycling 4-base Bsal-HFv2 37-16 °C cycling 4-base BsmBI-v2 42-16 °C cycling 4...enzymes and cycling conditions (see Table 1).   Enzyme Temperature Incubation time/cycling Overhang...MM 42-16 °C cycling 4-base BspQI, 1x NEBridge Ligase MM 42-16 °C cycling 3-base  PaqCl...Buffer 37-16 °C cycling 4-base PaqCl, 1x NEBridge Ligase MM 37-16 °C cycling 4-base Sapl...              37-16 °C cycling 3-base Sapl, 1x NEBridge Ligase MM 37-16 °C cycling 3-base Table...set with the Type IIS restriction enzyme and thermocycler protocol. It will also identify overhangs with... Bbsl-HF®, 1x NEBridge® Ligase MM 37-16 °C cycling 4-base Bsal-HFv2® 37 °C Static 4-base...

  6. Hot Plasmids - October 2020

    Type
    Blog Post
    ...cell cycle length by examining the ratio of blue to red cells. Cells undergoing rapid cell cycles have...C. elegans Fluorescent timers for studying cell cycle length The CRISPR corner New from the viral service....org/10.1101/2020.07.02.185249 Predicting cell cycle length with a histone-fused fluorescent timer By...By Kate Harten DeMaio Studies of the cell cycle attempt to understand development, regeneration, and certain...states. However, it’s difficult to observe the cell cycle in vivo, particularly if the cells are migratory...blue to red ratio, whereas cells with long or slow cycles are more red than blue (Eastman et al., 2020). ...expression of H2B-FT consistently predicted the cell cycle speed across these different cell types. In one ...

  7. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ... The Design-Build-Test-Learn cycle adapted for video making.   Cycle One: The Ironing Board Control...it was time to start our Design-Build-Test-Learn cycles. If you’re not familiar with this, it’s a method... WFH desk, and as a video control center.   Cycle 2: Zooming Forward Design: We produced a video celebrating...shot composition.  So back to the drawing board.  Cycle 3: A Return to HQ Figure 3: Even Blugene...in a different room.   Design: The previous cycle made me realize we needed to focus on ways to shoot... be in there with them. I knew from my previous cycles that I wanted to be able to talk to people and ...camera control, and extra hand sanitizer.    Cycle 4: Venturing into the Lab Design: The lab is the...

  8. Deep Dive: qPCR

    Type
    Blog Post
    ...molecule often work better for rapid-cycle PCR, such as the LightCycler, as it reduces the primer/probe actions...This separate thermocycle allows you to measure the intensity of signal over an entire cycle. Since signals...especially if you’re using a rapid-cycle machine such as a LightCycler, you’ll likely need to re-optimize...particular has been reported to work well in the Lightcycler while being relatively easy to multiplex (Thellwell...unless running a multiplex reaction or on a rapid cycle machine. Melt Curves An advantage of Sybr chemistry...paper for more information on optimizing for LightCycler machines.) Multiplexing It is possible to multiplex... IA; Choi JW;  Morlese J; Taylor G; Shaun S. LightCycler qPCR optimisation for low copy number target ...

  9. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    ...reaction cycle. However, it is common for dNTPs and other reagents to run low during the final cycles which...baths but is now easily done in a programmed thermocycler. For more details on this procedure check out...implies, analyzes the end product of PCR temperature cycling. The final PCR product is often visualized on a... use of a fluorometer feature built into the thermocycler to measure that fluorescent output. With this...concentration of the DNA during the PCR reaction cycles is continuously detected via a fluorescent signal...proportionally to the amount of product produced each cycle.  To determine the concentration of the starting...amplified DNA of a known starting concentration. The cycle in which the unknown DNA is detected compared to...

  10. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...integrase.  The IDLV life cycle When IDLVs are delivered to cells, the viral life cycle initially proceeds as...retroviridae, require integration for their life cycle to continue. They rely on a viral protein called...production (read our blog post on the retroviral life cycle for a more detailed description). Since the integrated...contain any of the viral packaging genes, the natural cycle of the lentiviral vector ends here and no viral ...Figure 1).  Figure 1: Comparison of the life cycle of a lentiviral vector with an integrase-deficient...expression diminishes as the episomes are lost through cycles of cell division, though the rate of loss greatly...Addgene blog Viral Vectors 101: The Retroviral Life Cycle Quick Guide to All Things Lentivirus More resources...

  11. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...its native context, the tetracycline repressor (TetR) can bind to the tetracycline operator sequences (TetO...preventing transcription. In the presence of tetracycline (Tet), TetR preferentially binds Tet over the...repressible system, Gossen and Bujard created the tetracycline-controlled transactivator (tTA). They fused ...TRE)), allowing transcription to proceed. When tetracycline or one of its derivatives is added, it binds...information on Addgene Tet plasmids, see our Tetracycline resource page. Cumate Switch Massie et. al developed...control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S...Signals FLEx Vectors Resources at Addgene.org Tetracycline Collection ...

  12. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    ...HDR events, ensure cells are actively cycling through the cell cycle stages. If cells are overly confluent...across cell types. Variance can be caused by cell cycle rates; rapidly diving cells will have higher HR ...operates predominantly in S and G2 phase of the cell cycle, when a sister chromatid is likely to exist as a...take this one step further, you can inhibit cell cycle regulatory proteins responsible for the transition...out of S phase, so cells stay in an HDR-promoted cycle longer. This approach may have some negative outcomes... if cells are arrested too long. Prolonged cell cycle arrest can lead to apoptosis in as little as 10 ...

  13. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    ...plastic and packaging material or by incorporating recycled paper and plastic. Aside from the more obvious...obvious swaps like using smaller bottles or recycled cardboard packaging, there are some less obvious improvements... Waste tubes are made of 100% post-consumer recycled plastic Reduced plastic in buffer bottles ... from packaging Made from 100% post-consumer recycled material Reduced cardboard packaging Plant-based... cardboard packaging by more than half 70% recycled materials Sustainable forestry certification Starch-based...Starch-based compostable bags for kit components Recyclable Yes, all kit components and packaging Yes...

  14. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    ...reverse tetracycline-controlled transactivator) is normally inactive and cannot bind the tetracycline response... inducible promoters. The positive inducible tetracycline ON (Tet-On) system, a versatile tool developed...response elements (TRE) in a promoter. Tetracycline and its derivatives serve as inducing agents to allow...Addgene.org: Inducible Promoters for Bacteria Tetracycline (Tet) On Expression Systems at Addgene Cre-lox...

  15. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    ... triggered to enter the lytic cycle.   During both of these life cycles bacterial DNA can be accidentally...inside, phages can follow one of two different life cycles: lytic or lysogenic. Lytic phages hijack the bacterial...that can infect other bacteria. In the lysogenic cycle, the phage’s genetic material is incorporated into...competence occur during the Helicobacter pylori growth cycle." FEMS microbiology letters 255.1 (2006): 148-155...

  16. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...by titration with different amounts of tetracycline. Tetracycline regulation is reversible (when the drug...with more elements of control. Tet promoters Tetracycline-dependent promoters are engineered by placing...constitutively silenced proteins. Addition of tetracycline or one of its analogs will induce these transcriptional...transcriptional level.     Fig. 2 Tetracycline on/off promoters and modes of action Check out...

  17. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...two PiggyBac integratable cassettes, both with doxycycline inducible promoters (Fig. 2a). The first cassette...contrast. In all the cell lines we tested, 1 µg/mL doxycycline was sufficient to saturate expression. When ...Next, perform a short (12-hour) induction with doxycycline prior to FACS, gating for the top quartile (or...producing cells. After FACS, return the cells to doxycycline-free media and expand the cells. Use a subset...engineered cells via regular intraperitoneal doxycycline injections to enable their tracking with ultrasound...

  18. Bringing Renewable Energy to Addgene

    Type
    Blog Post
    ...Xerox for recycle and reuse, reducing the need for the production of new plastic.  Recycling and reuse...areas provide multiple streams for composting and recycling in the event the object in question cannot be ... We participate in many styrofoam and plastic recycling programs in the lab as well. For a more in-depth...

  19. Bringing Sustainable Practices to the Lab: Innovations

    Type
    Blog Post
    ...our consumption beyond recycling.     Choosing more sustainable products Recycling is great for rerouting... post focused on streaming some lab waste for recycling- a process that’s integrated into waste management...Additional resources on the Addgene blog Learn about recycling initiatives for the lab Subscribe to the Addgene...

  20. Management for Scientists: Managing vs. Leading

    Type
    Blog Post
    ...workers into three cycles: Doers, Managers and Leaders. I like her use of the term “cycles” instead of levels...roles. At any stage in our careers we may be in a cycle of doing, managing or leading. All three roles are...need to do to be successful in a specific role or cycle. For example, if you try to lead when you are expected...

  21. Plasmids 101: Antibiotic Resistance Genes

    Type
    Blog Post
    ...permeability Bactericidal 10-100 ug/mL Tetracycline tetracyclin Binds 30S ribosomal subunit; inhibits...aliquots at -20oC and avoiding repeated freeze/thaw cycles will keep most antibiotics viable for at least ...in foil if a light-sensitive antibiotic like Tetracycline is used. Keep in mind that some E. coli strains...

  22. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ...2. Viral stocks may be sensitive to freeze thaw cycles. Depending on the researcher and the type of virus... of titer losses from 5% to 50% per freeze thaw cycle (Krajden et al., 1999;  Ugai et al., 2002). If the...However, polybrene is highly sensitive to freeze-thaw cycles and should be stored in single-use aliquots. Never...Krajden, Mel, et al. "Effect of multiple freeze-thaw cycles on hepatitis B virus DNA and hepatitis C virus ...

  23. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ... vector. In the absence of tetracycline or a derivative such as Doxycycline (Dox), the Tet-controlled ...needs to be expressed a certain point in the cell cycle, a transfer vector with an inducible promoter (see...used inducible lentiviral vector system is the tetracycline (Tet)-regulated system. You can choose either...

  24. Bringing Sustainable Practices to the Lab: Easy Action Items

    Type
    Blog Post
    ...other posts on lab sustainability, check out my Recycling and Innovations posts. Making small changes in...sustainability project. This could be: Setting up a recycling program at your institute. Set your -80 °C freezer...Additional resources on the Addgene blog Learn about recycling initiatives for the lab Subscribe to the Addgene...

  25. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    ...editing The best studied Cas12b nuclease from Alicyclobacillus acidoterrestris (AacCas12b) exhibits optimal...genome editing system that could be regulated by doxycycline. Now, they’ve developed another AAV toolkit -...system is regulated not only temporally with doxycycline, but it can also be regulated spatially with ...

  26. The Stingy Scientist: How the Baby Gel Box Was Born

    Type
    Blog Post
    ...creating less waste.  Most communities seem to have recycling programs and it seems to me that most Addgenies...fewer people use plastic bags.  We also try to recycle our discards according to the Cambridge, Massachusetts...chipped or imperfect equipment.  We can’t safely recycle these materials either.  Test tubes aren’t cheap...

  27. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    ...repeated cycles will favor accumulation of the latter products. As noted above, a single cycle of cleavage... Unlike HDR, NHEJ is active throughout the cell cycle and has a higher capacity for repair, as there is...

  28. Plasmids 101: Cre-lox

    Type
    Blog Post
    ...these components as part of its natural viral life-cycle, and researchers have adapted the components for...recombination” (although later referred to as the “cyclization recombinase”), is a 38 kDa protein responsible...the Cre inducible (such as with tamoxifen or doxycycline), the Cre recombinase can be expressed only in...

  29. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ...added to the mastermix that are read after each cycle. The raw data is then normalized to a reference ...dependent on number of samples rather than number of cycles.  Choosing a method  As with all experiments, a...analysis of copy number variants based on real-time LightCycler PCR. Curr Protoc Hum Genet. 2014;80:7.21.1-7.21...

  30. PhD Applications After COVID

    Type
    Blog Post
    ...there are many places where the PhD application cycle becomes much more complex. “How do I reach out to...respond to your email. During my PhD application cycle, I reached out to around twelve professors and heard...like that! For instance, during my application cycle, two schools on opposite sides of the country had...
Showing: 1 - 30 of 210 results