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...system. The first two can serve as controls in a variety of experiments, while rtTA can be used to activate...that can be used to deliver fluorescent proteins to cells as determined by the serotype. You can use these...that you can use to map neuronal connections. Expanding the viral vector knowledge base: You can help! We...from select vectors in the repository yourself, you can now order ready-to-use virus directly from Addgene...many researchers we’ve met with have asked if we can help them share and distribute other reagents. Virus...while still ordering viral vectors. Virus production can be a frustrating process that requires a large investment...viral production and testing so that researchers can use our educational resources and experience to their...

...factors that can affect DNA stability in solution are pH and DNase contamination.  Extreme pH can denature...sample over time and determine which experiments you can effectively use the DNA. pH and DNases affect DNA...stranded DNA are repaired by base excision repair, but can lead to mutations in single stranded DNA undergoing...undergoing replication.   DNase on the other hand can digest and degrade the double helix, fragmenting plasmid...degradation, and/or the use of chelating agents that can be introduced in the purification protocol to remove...substantial degradation and which experiments you can use your plasmid.  DNA elution choices: TE, Tris ...the stability of your DNA, it will limit what you can do with these samples. This is because divalent cations...

...tissue sample can also need significant optimization for the sample type and size.) It can be quite the...protein and DNA are no longer bound, and the DNA can then be isolated and studied through a variety of...application for exploring these interactions, and can give valuable insight into DNA expression and protein...approach for the faint of heart!  The Process ChIP can be done for either tissues or cells. The process ...most ChIP experiences, but the increased binding can mask the epitopes antibodies recognize. It is therefore...before moving on to the next step. Typically, this can be done by cutting the tissue sample up and then ...usually the preferred method, enzymatic digestion can be appropriate when your protein of interest is a...

... gradients can be informative, so spend some time thinking about which type of gradient can help answer... gels can overheat. Running your transfer in the cold room or packing the box with ice packs can help ...from the same lot as you need or can get from the manufacturer. You can also consider using monoclonal ... high, you can consider adding a detergent like Tween-20 to your washes — although this can be harsh and...Western blots can be tricky to get right. If you’ve thoughtfully done the technical design of your blot... optimization and troubleshooting.   Before you can optimize or troubleshoot a western, you’ll need to...Created with biorender.com   Alternatively, you can run a reagent gradient. In this approach, you’ll ...

...is stored in the cloud, so you can access it from anywhere, and it can be shared with others. The Web ...Connector, papers can be saved from Firefox or Chrome browsers with just one click. Papers can be cited in ...LibreOffice, or Google Docs. Files can be synced across devices, and you can share your library with others...calendar after completing your writing goal, which can serve as positive reinforcement for a habit. The ...technique timer  One time management technique that you can use as part of your writing schedule is the Pomodoro...is called a “Pomodoro.” After four Pomodoros, you can take a longer 15-30 minute break. The method is named...word suggestions. De-jargonizer The De-jargonizer can identify jargon in your writing. While it’s not possible...

...inactivating genes. CRISPR can target a genomic sequence and create a break that can be repaired by homologous...designed in the lab. Specially constructed plasmids can be used to replace genes in yeast, mice, or Drosophila...This final product of non-homologous recombination can survive positive selection, but it will be sensitive...selection marker. Since an antibiotic resistance gene can affect a cell's phenotype, researchers usually remove...through the insertion of flanking LoxP sequences, you can remove the gene through the addition of Cre recombinase...the neomycin resistance gene is floxed so that it can be removed post-selection through Cre-mediated recombination... (knocked in). As seen in the above example, you can remove a floxed resistance gene using Cre recombinase...

... equipment can be defined as equipment with capital costs greater than $5,000 but this can depend on the...where you can start to identify which items aren’t critical to your research success. If you can get away...loss of time, can exceed the cost of the new equipment. That’s why a resource budget can help you compare...your research, negotiations can be much easier. However, negotiations can still go horribly awry when ...resources needed to accomplish your science, and can include: equipment, reagents, animals, glassware,...emotional weight. Therefore, too much focus on money can dissuade a productive negotiation. A more productive...much those resources will cost and how these costs can be reduced. Negotiating is easier when both parties...

...targets by CRISPR can accelerate the drug pipeline. High-throughput CRISPR screens can be used to inhibit... may be most effective. Once drug candidates are isolated, they can be validated for specificity to their...or their direct target can be identified if it isn’t known. If a strong candidate emerges, downstream optimization...added QC is where CRISPR can save time and money to make sure quality candidates are selected.      ... targeted design. Alternatively, a disease model can be generated, and drugs screened for a change in ... workflow? CRISPR both speeds up the process and can help weed out a lot of poor drugs before time and... affect disease progression or onset. These hits can reveal individual genes or sometimes even genetic...

...Protein Oligomerizes in Cells GFP oligomerization can lead to misleading artefacts in your experiments....Medicine, to learn how problematic oligomerization can be and how to avoid it in your experiments. Better...proteins are incredibly useful, chemical fluorophores can still be used in a wide variety of experiments. Read...produce reporter proteins. These toehold switches can be used to quickly detect the presence of Zika virus...School of Medicine, this post describes a tool you can use to study basic virus biology while leaving virulent...transcription as well as reporter genes. To learn how you can use minigenomes in your research, check out this ...-bodies, “force generating protein machines that can extend and retract over many microns in response ...

...system or cell type can completely fail in a different system (Issa et al., 2023). This can be particularly..., which can lead to cells in far away brain regions expressing your gene of interest. This can cause issues...circuit can occur. Promoters The promoter controls how the gene of interest will be expressed; it can be ubiquitous... chicken-beta-actin or CAG can drive strong early gene expression, but can become methylated over time...tyrosine-hydroxylase (TH), can limit which cells express your gene. Off-target expression can still occur (Fig....Using AAV vectors in complex biological systems can be tricky at best, and downright infuriating at worst... validation first.  AAV-mediated gene expression can differ extensively depending on the experimental ...