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We narrowed to 1,052 results for: FER

Showing: 221 - 240 of 1052 results

  1. 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...

  2. 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...

  3. 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...

  4. 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...

  5. 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...

  6. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    ...an antibody but want to know if different combinations or even different chimeras might function better...antibody-based cancer therapeutics.    References and Resources References Chan, A. C., & Carter, P. J. (2010...functions, the constant regions can be altered for different isotypes, and so much more. How you engineer an... variable; sdAb: single-domain antibody (often referred to as nanobodies). Created with BioRender.com....specific goal, like improved specificity. There are different ways to achieve this goal, including error-prone...same general cyclical process (Figure 2). The differences typically come in when generating the library...that contains a variety of degenerate codons (different codon sequences that code for the same amino acid...

  7. 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...

  8. 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...

  9. 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...

  10. 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...

  11. 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...

  12. 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...

  13. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...24, 2025 by Alyssa Shepard.   References and Resources References Beumer, K. J., Trautman, J. K....Cong worked with Addgene Senior Scientist, Matt Ferenc, to organize the CRISPR forum's most popular questions...double nickase system is that you can express two different chimeric gRNAs with the Cas9 nickase which will... like the wildtype Cas9 does. There are a few references for the double nickase system, including one ... my target sequence? A3: The human U6 promoter prefers a 'G' at the transcription start site to have high...system probably have better specificity. A good reference is Ran et al. (2013). As for target selection...most guides would work, but they sometimes have different efficiency). And it is best if the cut site is...

  14. 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...

  15. 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...

  16. 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...

  17. Sequencing Primers

    Type
    Guide
    ... Plasmid Reference Molecular Biology Reference Sequencing Primers Sequencing Primers...convenience. For reference information, please consult Addgene's Molecular Biology Reference Page . All listed...AGTCAAGTAACAACCGCGA 3' end of luciferase, forward primer LucNrev CCTTATGCAGTTGCTCTCC 5' end of luciferase, reverse primer...sequences listed on the left are provided for your reference. Addgene does not distribute primers. For sequencing... primer LucNrev CCTTATGCAGTTGCTCTCC 5' end of luciferase, reverse primer M13 Reverse CAGGAAACAGCTATGAC...GGGCTGGCAAGCCACGTTTGGTG 3' end of glutathione-S-transferase, forward primer SP6 ATTTAGGTGACACTATAG SP6 promoter...GGGCTGGCAAGCCACGTTTGGTG 3' end of glutathione-S-transferase, forward primer pGP704-R AACAAGCCAGGGATGTAACG...

  18. 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...

  19. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...digestion be sure that the buffers and temperature are compatible between the different enzymes. Always check...ligation mix in different bacterial strains (stbl2, NEB stable, etc.), and/or use a different temperature ...want to assemble together in a combination of different ways according to your cloning strategy: Restriction...from other DNA fragments and you will remove the buffers used in the previous reactions. This will increase... There are multiple ways you can assemble the different parts of a plasmid based on the cloning strategy...attention in the areas of ligation between the different fragments If you don’t get any positive clone...might want to reconsider your approach, and try a different cloning strategy. Once you've identified potential...

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

    Type
    Blog Post
    ...current job situation. What can a coach do that is different from an adviser, boss or mentor? First, professional... transition out of the academic sphere into a different meaningful career in science. Supervisors in the...speaking at research institutions and at scientific conferences, I see that few scientists are aware of the benefits...few hours of coaching time can make an enormous difference so don’t assume that this will cost a fortune...successful outcome. How do I find a coach? Start with referrals. Are there scientists you know who have had a ...offices or professional organizations can provide referrals. There are bad coaches out there, so finding a...specializes in scientist coaching. Always ask for references and follow up with 3-4 to hear about those clients...
Showing: 221 - 240 of 1052 results