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 1,045 results for: fer

Showing: 221 - 240 of 1045 results

  1. Plasmids 101: Control Plasmids

    Type
    Blog Post
    ...multiple times and on different days. Using our example above, we would test different preparations of Plasmid...Plasmid A in different aliquots of cells on several different days. Biological replicates control for ...There are many, many different types of experiments carried out by scientists every day. Although the...the plasmid without the independent variable. Referring back to the experiment associated with Figure ...constitutively expresses a reporter protein (e.g., GFP or luciferase) that is either co-transfected with the test ... the results because it provides an important reference point when analyzing the specificity of the shRNA...variable should be consistent over time and from different preparations of the purified test plasmid expressing...

  2. Behind the scenes: Addgene’s new search engine and more

    Type
    Blog Post
    ...findability” of the information on the site. Different users have different strategies for finding what they need...the overall user experience of our website, the different website navigation paths, the organization and...homepage and navigational elements help users who prefer to browse and discover items in our catalog or ...resources through our main dropdown menu. For users who prefer to use search, our updated search engine is now...software team tailored Elasticsearch to support the different usage patterns the larger team identified. The...our catalog of materials and services is quite different from the way it searches through our educational...information in our search indices in order to support different kinds of matches: exact words, partial words, ...

  3. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...transgenes in iPSC-derived cells can interfere with their differentiation potential. Finally, retroviruses...reprogramming factors and wait for cells to de-differentiate. However it’s difficult, especially as a newbie...reprogramming proteins. Table 1: Key Features of Different Methods for Generating iPSCs. One important note...reactivation of Myc or other oncogenes after differentiation has been linked to tumor formation in iPSC-...drawback is that lentiviral vectors insert in different genetic locations in each transduced cell, so ...a single polycistronic transgene or with four different adenoviruses, each expressing one factor. These...mutagenesis, adenoviral vectors are considered a safer way to express reprogramming factors for therapeutic...

  4. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...components expressed from different plasmid backbones. They noted that backbone differences such as the order,...assess how different lentiviral plasmid backbone elements impact viral titer and to compare different Cas enzymes...Hurt for his expertise and input.  References and resources References Weber, Ernst, et al. “A Modular ...review common uses of MoClo and the kits Addgene offers to kickstart these applications, no matter what...product in an affordable way, researchers test many different combinations of promoters, terminators, solubility... excellent option (Iverson et al., 2016). CIDAR offers a one-pot reaction wherein multi-part assembly ...is at your service (Lee et al., 2015). This kit offers a broad range of parts, including copy number machinery...

  5. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...this FRET you speak of? Förster Resonance Energy Transfer (FRET) was originally described by Theodor Förster...alternative acronym Fluorescence Resonance Energy Transfer. Unlike the typical excitation and emission of...excited fluorophore, FRET involves a non-radiative transfer of energy (i.e. no emitted photons) from the excited...excitation by absorption of a photon 2. Energy transfer from the excited donor directly to the acceptor...the acceptor fluorophore. Otherwise the energy transferred from the donor will not be able to excite the...other and be oriented appropriately for energy transfer via dipole-dipole interaction. The efficiency ... typical FRET range, FRET efficiency is used to infer an interaction between the fluorophores and serves...

  6. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    ... and reference on how to design PCR primers.  Types of PCR Since the invention of PCR, different PCR methods...PCR, which is commonly referred to as simply PCR, is used for an array of different molecular biology techniques...Analysis RNA detection     References and Resources References Shen C-H (2019) Amplification of... methods have been developed for different scientific applications. These PCR methods all use the same...same basic PCR set up and steps but differ in how the PCR products are analyzed.   End point PCR End point...alleles or mutations. ddPCR also does not require a reference or standard curve, which can be time consuming...Multiple template PCR - multiple templates with different primer pairs that align to the target region of...

  7. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...updated by Ina Ersing in August 2023 References and resources  References  Aurnhammer, C., Haase, M., Muether...required adenoviral helper genes are supplied on two different packaging plasmids: the pHelper plasmid containing...expressing Ad E1A and E1B, can be transfected with your transfer plasmid, carrying your gene of interest, and the...virions out. We recommend using a Tris-based lysis buffer and sonication.  The obtained virions from cells...remove the density gradient material by dialysis/buffer exchange and further concentrate the virions if...titering assay, you’ll need to understand how different titers are measured. A TCID50 measurement reports...have a high-quality plasmid standard and rAAV reference material of a known titer on every qPCR run. Note...

  8. Zebrafish as a Model for Behavior: Swimming into the Optogenetic Spotlight

    Type
    Blog Post
    ...Because of the differing peak absorption wavelengths and light intensity requirements of different opsins, studies...behavioral studies because it is non-invasive and offers a high degree of control over time and space. Zebrafish...simultaneously and still be individually activated with different lights (Baier et al., 2009).  Zebrafish behaviors...assays can be done on zebrafish at 5 days post fertilization, including escape behavior and spontaneous swimming... do so, the lab expressed the LiGluR opsin in different subsets of spinal cord neurons using the Gal4/...Gal4/UAS system. This created fish lines with different groups of neurons that could be activated with light...spontaneous swim or escape, was elicited when these different neuron groups were activated or “turned on.” In...

  9. DNA Purification Without a Kit

    Type
    Blog Post
    ...could just make their own buffers when the kits buffers ran out. Homemade buffers performed the same as those...maxi, giga) which requires you to buy different kits for different purposes, with the Kim and Morrison ...Read our Molecular Biology Reference Visit our protocol video library References Kim, Y., & Morrison, S.... need them and to accumulate a bunch of unused buffers because you’ve run out of columns. Kit-less DNA...precipitated with alcohol and resuspended in TE buffer. After this, the DNA is ready for use! Plasmid ...released by dipping the cellulose directly into a PCR buffer, where DNA or RNA can be amplified. Compared to...times. DNA purification kits don’t come with enough buffer for this many uses of the columns, but the Zhao...

  10. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated...of cells the vector could infect. Finally, the transfer plasmid encoded the desired transgene flanked ...facilitates nuclear export of transcripts). A different viral envelope protein (Env), usually from VSV-G...packaging system Although the 2nd generation system is safer than the original system, the possibility of creating...-competent virus via recombination between the transfer and packaging plasmids does exist. To further ...created the 3rd generation system (Figure 3), which differs from the 2nd generation in two key ways. First,...containing a Tat-independent promoter. Although safer, this system may be more cumbersome to use and lead...

  11. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    ... probes, LDR can reliably detect single-base differences. The original creators of this technique visualized...fidelity mean for a ligase? DNA ligases generally prefer fully Watson-Crick base-paired dsDNA substrates.... The structural/mechanistic reason for this differential is not known for certain, but may have to do...This methodology has been used to rapidly screen buffer conditions for Taq DNA Ligase and their effect ...upstream probes and four down-stream probes, each differing only in the base at the ligation junction. Thus...Sixteen separate pools were prepared, each with a different complement strand covering all 16 possible NN ...each possible pairing resulted in a product of a different length, with products identifiable and quantifiable...

  12. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...when interpreting the outcome.    References and resources References Aschauer DF, Kreuz S, Rumpel S (2013...2, and 5 Display Differential Efficiency and Cell Tropism after Delivery to Different Regions of the Central...theoretically this strategy should work, these vectors suffered from high levels of spurious expression.  As such... recognized by Cre, but they are comprised of different sequences (loxp and lox2272) and can not recombine... flank a transgene (homologous pairs) or used different scrambled sequences to flank the transgenes (non-homologous...  The AAV serotype used can exhibit profound differences in tropism between cell types (astrocytes, microglia.... 2013, O’Carroll, SJ. 2021). This is due to differences in the expression of receptors, post-translational...

  13. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

    Type
    Blog Post
    ...codon usage biases of different organisms. Different species have different preferences for the codons they...the gene may not express well because of these different codon usage biases. But if a gene’s sequence was...could be determined through comparisons with a reference sequence database. Through a series of simulations...Kunjapur says. Percent sequence identity was a different story - the team found that the percent sequence...sequences is Proteobacteria - the most frequent transfer of genes is from Proteobacteria to mammalian expression...natural gene sequence is, the less likely it is transferred into another organism in the lab. Predicting ...and synthetic biology is expanding rapidly.  References 1. Kunjapur, Aditya M., Philipp Pfingstag, and...

  14. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ... C improved assay reliability. Different primers can have different annealing efficiencies (Wang et ...of samples from different sources should not be compared. If using AAV from different sources in the same...solution, and generally these methods measure different characteristics of the solution. These characteristics...because each sample with be quantified with a different primer pair. Ct values vary based on the amount...and low-concentration samples can also behave differently in PCR, even if both samples fall within the ...absolute quantification by using the universal AAV Reference Standard Material (AAV SFM), which is an AAV sample...addition to the AAV RSM, we include a second AAV reference sample of known titer, which is at a higher titer...

  15. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...plasmids are compatible, different CBPs bind to each plasmid type, and different NTPases separate the plasmid...plasmids at Addgene Read our Molecular Biology Reference References  del Solar G, Giraldo R, Ruiz-Echevarría...incompatibility is defined as the inability of different plasmids to be maintained in one bacterial cell...al., 1998).  In cases of compatible plasmids, different negative regulation systems control plasmid replication...regulation system cannot distinguish between the different plasmids. The concentration of negative regulation...diffusion) to make this happen.  There are several different versions of the plasmid partitioning system in...identification issue: the CBPs in the cell can’t tell the difference between plasmids if they have the same centromere-like...

  16. Career Insights: Technical Support Specialist

    Type
    Blog Post
    ...expect, different companies produce different products and therefore need people with different areas of...life science PhDs can offer: vast practical lab expertise, experience in different scientific fields, and... as: “Can I use this product under conditions different from those specified on its data sheet?” or “Do... questions customers ask, and there are also differences in customers’ personalities and their expectations...do jobs in technical support have? There are different names for technical support roles. Examples are...distinction between these job titles, they may be different regarding the levels of lab experience and scientific...communication skills by working with people from many different backgrounds and realizing that the same approach...

  17. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    ...variants differing in kinetics, affinity, and ∆F/F0. Introduce the two variants in three different membrane-targeting...fluorescent signals upon sensing glutamate in mouse and ferret models. Furthermore, they created three affinity...medium affinity for glutamate), that came in three different colors – blue (SF-Azurite-iGluSnFR), green (SF-iGluSnFR...glutamate signaling events, and accurately differentiating between synaptic vs. extrasynaptic glutamate...Figure 1   Increased versatility through two different variants The yellow (v82) and lime green (v857... Figure 2: Glutamate titration curves for the different iGluSnFR variants. Image adapted from Aggarwal...Figure 3: pH titration curves and ∆F/F0 for the different iGluSnFR variants. Image adapted from Aggarwal...

  18. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...can't distinguish the different proteins from each other. To distinguish different target proteins in your...target. By using a mix of different antibody-fluorophore combinations for different markers of interest, you...occur in combination with that reporter, you can infer its presence through detecting the reporter's fluorescence...a process called Fluorescence Resonance Energy Transfer (FRET).   Figure 1: Emission spectra and...commonly used fluorophores. A fluorophore's brightness refers to the signal strength of the emitted photons detected...a global maximum at around 560 nm, leading to different levels of brightness, depending on the laser light's...these aren't strict rules and in practice the differences in brightness levels are often not that great...

  19. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ... in a different city, you can still develop a successful FRET-based assay since energy transfer only occurs...what's it good for? Fluorescence resonance energy transfer (FRET) is a biophysical interaction between two...first fluorophore (the donor) is excited, it can transfer energy directly via resonance to the second fluorophore...targeting the donor and acceptor fluorophores to the different biomolecules you’d like to test for interaction...that the targeting specificity requirements are different for the donor and acceptor fluorophores. The donor...Å away from the donor doesn’t result in energy transfer and thus, your specific FRET signal is unaffected...unaffected  Due to these fundamental differences in targeting specificity for donor and acceptor fluorophores...

  20. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ...overexpress or knockdown a gene, you might need different plasmids. Harvard’s DRSC/TRiP Functional Genomics...varying expression levels of your transgene in different fly lines. Also, if there is a chance you need...transgenes in one fly, having the transgenes in different loci allows for genetic recombination so that ...for direct comparisons between lines expressing different variants of YFG because the transgenes are found...remember that you’ll need to integrate them all at different loci. Fortunately, there are many docking sites...docking site is located). This amounts to a small difference of two or three weeks. 3. How would you like ...yourself. Depending on your strategy and the services offered by the company, you might have to map into which...
Showing: 221 - 240 of 1045 results