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We narrowed to 881 results for: NSI

Showing: 101 - 150 of 881 results

  1. Distribution to Industry

    Type
    Collection
    ...Standards Kit Cloning Eric Cantor Kit for using comprehensive end-joining ligation fidelity and bias data ...

  2. p53 Pathway

    Type
    Collection
    ...E3 ubiquitin protein ligase PTEN Phosphatase and tensin homolog PUMA BCL2 binding component 3 Reprimo TP53...

  3. Zebrafish Plasmid Collection

    Type
    Collection
    ...Neurodegenerative Disorders - Nicholas Cole Lab. A kit consisting of plasmids with four promoters, six fluorophores...

  4. Control AAV Preps

    Type
    Collection
    ... found to match all requested criteria. Please consider using fewer filtering options. Browse all Control...

  5. Ras Pathway

    Type
    Collection
    ...non-catalytic subunit gamma PTEN Phosphatase and tensin homolog RAC RAC1 RAC2 RAC3 Ras-related C3 botulinum...

  6. CRISPR References and Information

    Type
    Collection
    ...entries describing enzyme activity, experimental considerations, and more. CasPEDIA is a community resource...

  7. Optogenetics AAV Preps

    Type
    Collection
    ... found to match all requested criteria. Please consider using fewer filtering options. Browse all Optogenetics...

  8. Retrograde AAV viral preps

    Type
    Collection
    ... found to match all requested criteria. Please consider using fewer filtering options. Browse all Retrograde...

  9. Caltech Systemic Capsids

    Type
    Collection
    ...do you want to target CNS or PNS? Then consider species, and then cell type. For CNS: For infant...

  10. Biosensor AAV Preps

    Type
    Collection
    ... found to match all requested criteria. Please consider using fewer filtering options. *AAVrg = retrograde...

  11. Validated gRNA Sequences

    Type
    Collection
    ... Keep in mind that additional factors must be considered before using a validated gRNA sequence in your...

  13. Antibody Guide

    Type
    Guide
    ...Figure 1. The two arms of the Y structure are responsible for binding the antigen, and are called the antigen...isotype of the antibody. B cells, the immune cell responsible for producing antibodies, can change the constant...binding site. In B cells, the genetic region responsible for the variable region goes through a randomized...valency of 2 IgE Protects against parasites and is responsible for driving allergic reactions such as anaphylactic...requirements, monoclonal antibodies are typically more expensive than polyclonal antibodies. Environmental factors... Minibodies are engineered antibody fragments consisting of two scFvs bound to a CH3 domain. Fab fragments... protein. Fluorophores Fluorophores are light-sensitive molecules which emit a specific wavelength of ...

  14. Plan Your Experiment

    Type
    Guide
    ...Manipulation Application Cas Enzyme Additional Considerations Browse Plasmids Knockout Permanently disrupt...essentially function as a “knockout,” but be sure to consider potential consequences of having a non-functional...be no PAM site close to your desired edit site. Consider alternative Cas9 enzymes with different PAM sequences...completely avoid them. To increase specificity, consider using an engineered Cas enzyme with higher fidelity...to off-target activity, it is also important to consider factors that maximize cleavage of the desired ...with BioRender.com. When choosing your system, consider which Cas enzyme you are using (as species and...for your experiment) and the expression time (transient or stable). For plasmid and viral-based approaches...

  15. Optogenetics Guide

    Type
    Guide
    ... the following enhancements: Increased light sensitivity Example: ArchT Increased photocurrent amplitude... Volvox carteri 570 Chronos High-speed, light-sensitive channel from Stigeoclonium helveticum 500 Chrimson...from Halorubrum strain TP009. Improved light sensitivity over Arch. 566 eArchT 3.0 ArchT with addition... delivery system. Here are some key factors to consider: Optogenetic excitation or optogenetic inhibition...neural populations at distinct times. Temporal considerations. Temporal precision is key in optogenetic experiments...Rhodopseudomonas palustris bacteria. Key feature is the sensitivity to 740- to 780-nm near-infrared light. 740–780...488 LOV domains LOV-based Light-oxygen-voltage-sensing (LOV) domains are protein sensors used by many ...

  16. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...the viral genetic material inside. The AAV genome is ~4.7 kb long and consists of two open reading frames...ITRs is limited by the physical space available inside of the small AAV capsid and directly correlates...Genome Integration Recombinant AAVs are generally considered to not integrate into the host genome. When wild-type...during production, as it carries the Cap region responsible for the production of capsid proteins. AAV2 is...it was the first to be well-characterized and extensively studied, and has a broad tissue tropism. Table...information. Self-complementary AAV (scAAV) One downside of the use of AAVs as viral vectors is their single-stranded...common points of entry. Biosafety should always be considered with respect to the precise nature of experiments...

  17. Chemogenetics Guide

    Type
    Guide
    ...molecule ligand. The ideal chemogenetic tools are unresponsive to native ligands and are engineered to respond... development of mutated receptors that while unresponsive to endogenous ligands, could be activated by... coupled receptors, but unlike RASSLs, show insensitivity to endogenous ligands, have low constitutive...proteins of a light-emitting luciferase and a light-sensing optogenetic element. When the luciferase substrate...luciferase-fluorescent protein fusions, to more-sensitive opsins has led to improved LMOs with increased...chemogenetics experiment, here are some things to consider: Chemogenetic activation or inhibition. Do you... a study in rats using an hM4Di receptor might consider an alternative DREADD ligand such as DCZ versus...

  18. CRISPR Guide

    Type
    Guide
    ...fusing the anti-CRISPR protein AcrIIA4 with the photosensitive LOV2 domain . Several labs have also controlled...There are two main types of base edits: Base transitions - edit to same classification of nucleotide base...pyrimidine-to-pyrimidine), which catalyze base transitions. Cytosine base editors are created by fusing ...integrated along with the sequence, so the edit is not considered “scarless”. Figure 7: Overview of CRISPR transposases... target gene function. It has also been used extensively to screen for novel genes that regulate known...simplest dCas9-based activators and repressors consist of dCas9 fused directly to a single transcriptional...silencing across much of the human genome following transient expression of the dCas9 fusion, with the epigenetic...

  19. Lentiviral Vector Guide

    Type
    Guide
    ...plasmids (mostly third-generation) can be used in transient transfections, but expression will be greatly ...plasmids cannot be used in direct transfections for transient expression of a transgene. For more detailed protocols...stable expression and wide tropism. Addgene has an extensive collection of lentiviral CRISPR libraries, including...initial infection. Third-generation systems are considered safer than second-generation systems because ...transfer plasmid is an oncogene. Additional consideration for oncogenes should be made on a case-by-case...common points of entry. Biosafety should always be considered with respect to the precise nature of experiments...9765382 Durand, S., & Cimarelli, A. (2011). The inside out of lentiviral vectors. Viruses , 3 (2), 132...

  20. Adenovirus Guide

    Type
    Guide
    ...a protein capsid and the viral genetic material inside. Adenoviral genomes range between 26–45 kb in length...ITRs is limited by the physical space available inside of the adenoviral capsid and directly correlates...used generation for research purposes. The system consists of two plasmids that eventually recombine to form...expression delivered through rAdV vectors is normally transient and short-lived. Second- and third-generation ...common points of entry. Biosafety should always be considered with respect to the precise nature of experiments...the Adenoviridae family. Episome Genetic element inside the cell that remains as a part of the eukaryotic...

  21. Promoters

    Type
    Guide
    ...equivalent called the Pribnow box which usually consists of the six nucleotides, TATAAT. Proximal Promoter...position, simply titled the -35 element, typically consists of the sequence TTGACA and this element controls...have also been discovered in eukaryotes. Operons consist of a promoter, which is recognized by the RNA polymerase...transcribes messenger RNA (mRNA) which is the RNA responsible for providing a stable template for the translation... RNAs (tRNA), the adaptor molecules that are responsible for bringing amino acids to the ribosome when...

  22. Modular Cloning Guide

    Type
    Guide
    ...gene-editing tools, and more. The MoClo system consists of three sets of cloning vectors (Level 0, 1, ... for a variety of applications. EcoFlex MoClo Extension, Volume I Bacterial Expression Mikhail Shapiro...and expression tuning in E. coli . CIDAR MoClo Extension, Volume I Bacterial Expression Richard Murray ...used with plasmids from the MoClo-YTK . EMMA: Extensible Mammalian Modular Assembly Toolkit Mammalian ... to generate mammalian expression vectors for transient or stable transfections. PUF Assembly Kit Mammalian...

  23. Technical Design of a Western Blot

    Type
    Blog Post
    ...chemicals, and are not always consistent. Precast gels, while expensive, reduce protocol time, can be ...chemical-based blockers, while much more expensive, are consistent and stable at room temperature for 1–2... Direct detection method Less sensitive; usually more expensive Nitrocellulose or low-fluorescence...Multiplex Fluorescent proteins Less sensitive; more expensive Strip/Reprobe PVDF membrane Increased...specificity, lower cross-reactivity, and are more consistent lot-to-lot. The highest reproducibility comes...quantitative assays. On the other hand, they are not as sensitive to lower amounts of protein, and can require more...  Lysis buffer When choosing your lysis buffer, consider the subcellular location of your protein. Proteins...

  24. RANbodies: Reporter Nanobody Fusions

    Type
    Blog Post
    ...antibody is less sensitive than HRP RANbody detection, it is approximately as sensitive as detecting the...detection is less sensitive than HRP RANbody detection but approximately as sensitive as antigen detection...approximately as sensitive as detecting antigen with a standard primary/labeled secondary Less sensitive than HRP-RANbody...approximately as sensitive as detecting antigen with standard primary/labeled secondary   Less sensitive than HRP-RANbody...antibody and a ≥100-fold increase in signal over intrinsic fluorescence. Increased signal is likely due to...secondary antibody. Chicken IgY This RANbody reporter consists of two chicken IgY domains. Chicken IgY domains...detected enzymatically with HRP substrate More sensitive than detecting antigen with a primary/ labeled...

  25. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...      Figure 1: A protein gradient. The density of the bands, which increase as protein concentration... buffers and strengths. If that’s not working, consider using a subcellular fractionation kit or a lysing...re seeing protein aggregation, you may want to consider changing your incubation temperature. Some proteins...aggregate at 95 °C, a common lysis temperature, so consider a longer incubation (10–20 minutes) at 70 °C, ...If you need very accurate protein measurements, consider using an unstained protein ladder, which will ...ladder on the lefthand side. Whatever you do, be consistent and document it.   Did it work? Save yourself...reversible staining method of choice is appropriately sensitive. If using a PVDF membrane, you can dry the membrane...

  26. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...detailed below. Voltage Sensitive Domain (VSD) GEVIs Simply put, VSD-based sensors consist of a VSD fused to...gated ion channels all depend on transmitting and sensing electrical potential. Thankfully, there is a tool... These tools have two main elements: a voltage-sensing component and a fluorescent output. The GEVI family...divided up structurally into 2 main groups: voltage sensitive domains and Rhodopsin containing GEVIs. Chemigenetic...fluorescent reporter and the mechanism of their voltage sensing component, which separates them into two distinct...indicator, which can make the kinetics of voltage sensing to signal output slightly slower than other approaches...approaches. Functionally, the voltage sensitivity of the available VSDs has been successful even at the...

  27. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...Specific ​H​igh ​s​ensitivity ​E​nzymatic ​R​eporter un​LOCK​ing) technique Sensitivity: Single molecule...Two-step-SHERLOCK has a sensitivity in the zeptomolar range while One-step-SHERLOCK has a sensitivity in the femto-aptomolar...for rapid, ultrasensitive and visual detection of novel coronavirus SARS-CoV-2 Sensitivity: 4.6-11 copies...SARS-CoV-2 using a CRISPR based DETECTR assay Sensitivity:10-100 copies of RNA per microliter Time: ~40...two crRNAs that bind to two regions improves sensitivity of the assay. This reaction also contains SSB...one-temperature reaction, and is nearly single-molecule sensitive. In the case of HIV-1 and SARS-CoV-2 detection...detection of SARS-CoV-2 genetic material (CREST) Sensitivity: 10 copies of RNA per microliter Time: Not mentioned...

  28. Antibodies 101: Normalization and Loading Controls for Western Blots

    Type
    Blog Post
    ...the loading control: the intensity of each POI band is divided by the intensity of the loading control ...band intensity measurements become extremely susceptible to error, since tiny changes in intensity can ...membrane, which would be directly reported by band intensity. Researchers use later steps in this chain (blue...trust Assumption #1, any change in the normalized intensity of your POI could reflect either a genuine change...relationship between protein abundance and band intensity is an S-shaped curve. Sparse proteins may not ...problems for proteins of very different abundance, consider the following example gel. Figure 5: An... on the plot of protein abundance versus band intensity (right). Created with BioRender.com. On this...

  29. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    ...conditions. The use of low intensity light solves each of these problems; inexpensive tools such as LEDs can...precise, consistent, and controllable pulses of light. Complicated patterns of light intensity and wavelength... low-intensity light, so off-target effects should be limited. Phytochromes are light-responsive systems...Tabor have been working on making E. coli light responsive since 2005, when Tabor was working in Voigt's... heading of two-component systems (TCSs). TCSs consist of a histidine kinase that phosphorylates a response...Christopher Voigt’s lab created the first E. coli light-sensitive two-component system (TCS). Levskaya et al. fused...Tabor et al. subsequently engineered a second photosensitive TCS activated by green light; these two systems...

  30. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ...published Aug 30, 2018 and updated April 16, 2020. Sensitive and specific nucleic acid detection is crucial...nucleic acid detection however, either lack the sensitivity or the specificity to detect nucleic acids at...at low concentrations and/or are too expensive, time-consuming, and complex to use outside of standard...and Cas12a, to develop simple, portable, and inexpensive platforms to reliably detect nucleic acids at...and optimize the method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK and SHERLOCKv2...CRISPR-based diagnostics. SHERLOCK- Specific High Sensitivity Enzymatic Reporter UnLOCKING Cas13 (C2c2) was... your targeted nucleic acid. To increase the sensitivity of the assay, targeted DNA or RNA from a sample...

  31. Antibodies 101: The Four ELISAs and When to Use Them

    Type
    Blog Post
    ...are not as sensitive, however, so those studying low abundance antigens may want to consider alternative... ELISAs for their accuracy, specificity, and sensitivity (a good ELISA can detect picogram quantities ...advantages and disadvantages of each, and some considerations when using the method. ELISA basics All ELISAs...detectable product. Scientists compare the signal intensity of the sample to that of a standard curve to extrapolate... themselves, the process can be laborious and expensive. In some cases, conjugation methods can interfere...Incorporating a secondary antibody increases the sensitivity of the assay, since multiple secondary antibodies...sandwich ELISAs, in a competitive ELISA the signal intensity is inversely proportional to the amount of antigen...

  32. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ...retain fluorescent intensity during exposure to light, is another factor to consider. If your application...interiors to many inquisitive scientists. Seeing inside these specimens has offered illumination on biochemical...will outline some fundamental characteristics to consider when selecting a dye or ligand to make your selection...proteins would be SNAP-tag® and HaloTag®. These consist of a genetically encoded enzyme variant tag that...is bound through green light emission. Key considerations in selecting fluorescent dyes/ligands Effects...-conjugation affects protein function and its propensity to bind cellular components. Some dyes target...stably combined with nanoparticles, maintaining intensity at ~85% of its original value after 60 minutes...

  33. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...subcloning, expansion, and confirmation. This procedure can take several months but should be considered in labs...benefits and drawbacks of different methods (for comprehensive protocols for all of the methods discussed here...results in less than a day, but tends to be less expensive than an ELISA kit. In this approach, viral RNA...lentiviruses in parallel, this approach should be considered. Similar to the p24 assay, titering via measurement...When using qPCR based methods for viral titration consider using a reverse transcriptase mutant in a separate...carryover. Functional titer Functional titer is considered a more accurate method to quantify virus as it...of this approach is that it can be quite labor intensive. Target cells are transduced with serial dilutions...

  34. Molecular Biology Reference

    Type
    Guide
    ...additional strain-related information and a more extensive strain list. Strain Vendor(s) Genotype BL21 Invitrogen...sequencing chromatogram which provides the color and intensity of each fluorescent signal. Sanger can sequence...and C always pairs with G. RNA on the other hand consists of Adenine, Cytosine, Guanine and Uracil (U). ...

  35. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ...the least expensive of all the options discussed as it only requires PCR and the inexpensive DpnI enzyme...here to help with that! Four important factors to consider for any experiment are: Speed Cost Difficulty...For cloning in particular, another important consideration is: The number of inserts ...as some methods...the following cloning methodologies have been extensively described in other articles; so if you are interested...little as 30 minutes! Alternatively, you might consider Gibson cloning. In this technique, you first perform...order determined by their homology. You might also consider FastCloning, first described by Chaokun Li and...Finally, TOPO cloning is another good option to consider when time is of the essence. This method requires...

  36. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    ...cellular-level events and behavior. It’s a problem that he considers “one of the greatest challenges of neuroscience...particular advantage for observing events taking place inside cells that are on the move, such as T lymphocytes... calcium-binding sites as possible for added sensitivity. As reported in Nature Methods, the FRET changes...essentially all of the standard organisms for which extensive genetic tools are available. While he recognizes... he said. “It gives you more information than intensity-based readouts.” Griesbeck also points out that...approach he and his colleagues took to optimize sensitivities can now be applied to dozens of other published...for visualizing the transcription factor cAMP-responsive element-binding protein (CREB) in living cells...

  37. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ... One consists of NLuc fused to enhanced green fluorescent protein (GpNluc) and the other consists of NLuc... (FRET) probes. These probes generally consist of a responsive protein (e.g. calcium binding protein) ...membrane-inserted voltage sensing domain (LOTUS V; Luminescent Optical Tool for Universal Sensing of Voltage). With...how bright you can make a reaction but for how sensitive you can make a reaction. NLuc requires 100-fold...) fused to two fluorescent proteins. When the responsive protein binds to calcium it changes conformation.... In well-designed FRET-based biosensors, the intensity of the FRET signal can be correlated to specific...No photoexcitation High Ion specificity High sensitivity Easy expression Gives ratiometric data The result...

  38. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...classes of optogenetic tools, and what factors to consider when choosing one.     Figure 1: Example...or inhibit specific neurons without needing to consider secondary signaling pathways. There are many different...different opsin (natural and engineered proteins responsive to light in combination with retinal) variations...reviewed in Abreu & Levitz, 2020), including light-sensitive chimeric GPCRS (OptoXRs) (Tichy et al., 2019),... the extracellular domain of endogenous light-sensitive receptors, like mammalian rhodopsin, and the intracellular...Created with BioRender.com.   Important considerations for using optogenetic tools Once you’ve selected...specific tool. There are many, many options to consider, and the following factors will help you determine...

  39. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    ...your plasmid yield is sub-optimal? If you are consistently getting sub-optimal plasmid yields from your...Addgene uses and recommends DH5𝝰! This strain consistently gives good, high quality preps due to its endA1... this handy guide from OpenWetWare for a more extensive listing of common E. coli strains. Increase the...media or adding supplements can improve your cell density, leading to an improved plasmid DNA yield. Culture...supplements to the media You can achieve higher cell density using a nutrient-rich broth such as Terrific Broth...in moderation) may also serve to increase cell density and plasmid yield. Start with a single, fresh colony...Increasing the incubation time can improve the cell density and therefore the plasmid yield. You can optimize...

  40. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...each event to a channel number based on intensity. Higher intensity events are assigned to higher channels...a user will compare the shifting intensity of events. Since intensity positively correlates with expression...cabinet with cloning rings or following labor-intensive limiting dilution protocols? If this sounds like... the cell’s size, complexity and fluorescence intensity.   What flow cytometers detect Cell size and... a high degree of internal complexity such as extensive membranes produces a greater SSC.  Protein expression...The electrical system converts the fluorescence intensity to a voltage pulse, called an event, and assigns...reporter on the N- or C-terminus of the protein. One downside of fusion proteins is that the tag can alter the...

  41. Antibodies 101: Buffers, Storage, and Conjugates

    Type
    Blog Post
    ... to be considered as well. In this post, we’ll touch on antibody storage, buffer considerations, and give...records, and consider the following questions as you move forward:  Was a light-sensitive conjugate on...you’re planning to run with the antibodies time-sensitive, using precious samples, or being compared to ...data in a manner that demands high accuracy and consistency?  If the answer to any of these questions is...saving the poor, abandoned antibody for a…less sensitive moment.   Fig 1: Blugene fading into darkness...fix it. HRP is easy to find, simple to use, not sensitive to light in its inactive state, and cheap. Unless... panel in the machine. Fluorophores are light sensitive, so antibodies conjugated to them need to be stored...

  42. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...more sensitive channelrhodopsin actuator for triggering action potentials at low light intensities, the...cumbersome. To overcome many of the limitations intrinsic to DNA-guided systems such as TAL effectors or...antibodies fused to sfGFP. This system amplifies the intensity of fluorescence signal and enables tracking of... H+ specific response cytoplasmic retention insensitivity to other ions In BRET, A mutant Renilla luciferase...dimers by fusing the SsrA peptide with the light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2...activator protein to drive transcription. The anchor consists of dCas9 fused to CIB1 and is targeted to specific... a photoswitchable fluorescent protein which transitions from emitting orange (λ= 565nm) to far-red (λ...

  43. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    ...fluorescence intensity remains. The first issue is that, in several cases, the fluorescence intensity decay ..., fluorescent proteins may display reversible intensity changes (Shaner et al, 2008; Bindels et al, 2017...experiment is performed that measures fluorescence intensity over time. In order to predict how fluorophores...proteins and by comparing how the fluorescence intensity changes over time, the photostability of the fluorescent...photobleaching rate does not depend on fluorescence intensity or protein distribution. Therefore, photostability...analogous to radioactive decay). Consequently, the intensity decay is often described by the t½, which is the... it is necessary to know how the fluorescence intensity evolves over time. The second issue is that, in...

  44. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    ...help with transitioning out of academia (because so many scientists are making this transition). Another...have experience and knowledge to help scientists transition out of the academic sphere into a different meaningful...not always as effective in mentoring for this transition. Second, a coach will help you set goals and ...successful outcome and the good ones take this responsibility seriously. As I make my rounds speaking at ...However most coaches who focus on scientists are sensitive to this and have coaching plans that are financially...here are a number I have come across for you to consider. Feel free to add other suggestions in the comments...scientific backgrounds. For a stated purpose of transitioning a scientist from academia to industry, you might...

  45. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ...2024.12.20.629623.  A fast and responsive voltage indicator with enhanced sensitivity for unitary synaptic events.... Z. (2024). A fast and responsive voltage indicator with enhanced sensitivity for unitary synaptic events... tags for single-molecule biophysics Fast and responsive voltage indicator ONE-GO biosensors kit Antibodies...collaborators recently created ASAP5, a faster and more responsive genetically-encoded voltage indicator (GEVI) ...ASAP5 selected for its improved kinetics and responsivity in screening ASAP3 variants. B) Electrical and...regimes with fast activation kinetics and high responsivity near the resting membrane potential, ASAP5 is...the formation of Gα-GTP by BRET. The biosensor consists of a YFP-tagged G protein and a NanoLuc® (Nluc...

  46. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ... problem. It is estimated that mycoplasma is responsible for up to 60% of the cell culture contamination...contamination (Uphoff, 2002). Mycoplasmas are considered to be one of the simplest and smallest bacteria. The absence... source of culture is irreplaceable or highly expensive). Detecting mycoplasma contamination Detection...Drexler, 2011). Culturing on agar plates/broth is considered the ‘gold standard’ for detecting mycoplasmas...noticeable color change. This method has high sensitivity but low specificity (Degeling, 2012). Bioluminescence...result interpretation might be difficult. The low density of mycoplasmas, contamination by other bacteria...you keep all the required equipment and reagents inside the laminar airflow before and while you are working...

  47. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...Gervais C, Grazzini E, Durocher Y. Transient Gene Expression in Suspension HEK293-EBNA1 Cells. Methods Mol...based on magnetic beads from very high cell density suspension. Biotechnol Bioeng. 2021 Sep;118(9):3499-...At its core, harvesting recombinant antibodies consists of separating antibodies from media, cells, and...this method, beads are incubated with the cell suspension. Antibodies bind ligands on the beads and a magnet...alongside your ladder and a protein standard. The density of the antibody bands can be measured against the...multiple contaminating bands or a few strong ones, consider increasing the number of washes performed during...conformational changes in the antigen or lower detection sensitivity of the assay. Where possible, it can be useful...

  48. CRISPR Antimicrobials

    Type
    Blog Post
    ...system both favors antibiotic sensitive bacteria and prevents sensitive bacteria from acquiring resistance...system. These CRISPR antimicrobials successfully resensitized a beta-lactam resistant population to the antibiotic... may result in plasmid loss and antibiotic resensitization, or may also cause cell death. Image from Beisel...et al. designed a two-phage CRISPR system to resensitize antibiotic-resistant bacteria (Figure 2). The...Bacteria that receive this phage are now antibiotic sensitive, but lytic phage-resistant. When the lytic phage...killed, antibiotics can be used to target the sensitive population. Yosef et al.’s proof-of-concept work....   Figure 2: Two-phage strategies to resensitize bacteria to antibiotics. A lysogenic phage infects...

  49. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...in yeast with Rosella, a pH-sensitive fluorescent biosensor A pH-sensitive fluorescent biosensor-based...-emission biosensor Rosella consists of DsRed connected to a pH-sensitive variant of GFP (SEP) by a 9 ...Virus (WNV) induced cell death. The Wu lab library consists of 77,406 individual gRNAs targeting a total of...Moduclar Cloning (CIDAR MoClo) Parts Kit. This Kit consists of a variety of plasmids containing many different...plasmids to the scientific community. This collection consists of plasmids containing genes from a crowd-sourced...scientists use GMAP, there will be a continuous expansion of compatible backbones, promoters, and genes ...all under the control of the cI857 temperature-sensitive repressor. The temperature-controlled expression...

  50. Hot Plasmids - November 2023

    Type
    Blog Post
    ...also incorporated LOV-Turbo into TransitID experiments.  Find TransitID plasmids here! These two developments...genetic cargo Optogenetic proximity labeling and TransitID Tracking cellular history with protein ticker ...37612276.   Optogenetic proximity labeling and TransitID By Mike Lacy Proximity labeling can identify protein...utility of this technique.  By engineering a light-sensitive LOV domain into the TurboID enzyme, Lee and Cheah...plasmids here! Next, Qin and Cheah et al. described TransitID, where two proximity labeling enzymes, TurboID...catalyzing the biotinylation of nearby proteins. B) TransitID uses sequential labeling by TurboID and APEX2 ...trafficking within and between living cells by TransitID. Cell, 186(15), 3307–3324.e30. doi: 10.1016/j....
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