...plants. Further opportunities with pSiM24 While these properties of pSiM24 are suitable for many applications... of DNA that will be transferred to the plant, similar to viral vectors’ terminal repeats. It’s a lot ...larger cloning capacity would be very useful: enter pSIM24! To build a better binary vector, Sahoo et al. ... Sahoo et al. demonstrate that this new vector, pSiM24, can also be used with biolistic methods, making...Agrobacterium infection. At approximately 7 kb, pSiM24 is 2 and 6 kb smaller than commercially available...blotting. Given its versatility and customizability, pSiM24 can be used not only for plant engineering and ... Kumar, Nrisingha Dey, and Indu Bhushan Maiti. "pSiM24 is a novel versatile gene expression vector for...

...can be used in combination to simultaneously perform modifications, simply by adding all the necessary ...allow any cloning procedure to be performed using a simple two-step, 2 hr protocol prior to transformation...linear fragments need to get into the same cell. For simple cloning requiring just one or two recombination...Subcloning - To move a gene from one vector to another simply amplify the vector and insert with separate primer...colonies. We routinely screen just two colonies for simple cloning procedures. Use high fidelity DNA polymerase...

... cloning techniques with higher efficiency and simpler procedures have been developed. Some of these include... excessive time requirements. FastCloning is a simpler yet reliable cloning technique that was developed...2. Li, Chaokun, et al. "FastCloning: a highly simplified, purification-free, sequence-and ligation-independent...

...treatment can photoconvert GFP from green to RED! This simple technique has been shown to work in plant, Drosophila...

...can bring down the morale of a group with their pessimism, Know-it all experts, who think their way is...

... have no fear - I’m here to introduce you to a simpler, yet more effective, CRISPR-based epigenetic approach...Sapozhnikov and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation, leaving...blasticidin or puromycin resistance genes.  It’s that simple! Good luck and happy editing!    Thank you to...

.... R., Abdul Aziz, A. T., Shueb, R. H., Zaid, M., Sim, B., Mustafa, M., Nik Yusof, N. K., Lee, C. K., Abu...

...., Rodriguez-Rozada, S., Dieter, A., Pieper, F., Sims, R., Tenedini, F., Bergs, A. C. F., Bendifallah,...cascade of the intracellular part of the receptor, similar to what the endogenous binding partner would do...over when and where the receptor is activated.  Similarly, PTLs are special, covalently linked molecules... light exposure to activate them. PORTLs use a similar concept, except the ligand is attached to a protein...

...Super-resolution techniques such as Structured Illumination (SIM), Single Molecule Localization (PALM/STORM), or Reversible... the world around us. Without contrast, objects simply vanish into noise. Due to its unrivaled contrast...Fluorescent Widefield Microscopy is the most common and simplest form of fluorescent microscopy. Collimated (non-converging...thicker three dimensional sample (see Figure 1); similar to an MRI or CT scanner. This is accomplished by...penetrates deeper into biological tissue. To prove this, simply hold a flash light to the palm of your hand. Although... excited at one time, limiting photobleaching. Similar to widefield, the entire field of view is excited...Website Other major microscopy manufacturers have similar information References 1. Lichtman JW, Conchello...

...Yuan, Y., Clark, P. M., Wang, Q., Martino, R. A., Sims, J. J., Bell, P., Raymond, A., Stanford, W. L., ... capsids cross the BBB via interactions with a similar protein called LY6C1. This protein is expressed...capsids were deposited by the Gradinaru lab. For simplicity, in each section we’ll list the Gradinaru capsids...Jang et al., 2023; Brown et al., 2021.) It has a similar transduction rate to AAV-PHP.eB in the brain, and...like the rhesus macaque, are approximately more similar to humans than New World Monkeys and rodents. Image...marmosets (Chuapoco et al., 2023). AAV.CAP-B10 shows similar behavior in marmosets as in mice, although the ... thymus, and transduce the heart and spleen at similar efficacy levels to AAV9 (Stanton et al., 2023)....

...decide to do simultaneous IF instead (Figure 1B). To label all your targets in one step, you simply use a mixture... each target. Alternatively, you can perform simultaneous detection with a mixture of primary antibodies...appropriate secondary antibodies. The general process is similar to standard immunofluorescence, but with some extra...antibodies before labeling the next target. B) Simultaneous Multiplex IF, where all the primary antibodies... Brewer et al, 2022 for a sample protocol.)  Simultaneous IF Depending on your needs, your sample, and...consideration when deciding whether you can do simultaneous or sequential labeling is what conjugated secondary...with Alexa Fluor® 488) then you can’t use them simultaneously because you won’t know which label the signal...

...recapitulate in vivo conditions similar to humans. For example, due to their simplicity, yeast have become an important...often as every three weeks so scientists can simultaneously observe several generations of mice at a time... mice are more genetically and physiologically similar to humans than many other model organisms, they...study human diseases. Diseases in mice show many similar phenotypes to human diseases and have successfully...Yeast (Saccharomyces cerevisiae) Yeast, one of the simplest eukaryotic organism, is a commonly used model ...in breads and other baked goods! Yeast is cheap, simple and easy to work with as they can survive in a ...scientists to easily manipulate them. Scientists can simply inject one-celled embryos with DNA or RNA to edit...

...standardized constructs and tools for the rational and simple shuffling of DNA regions. Golden Gate Assembly ...Assembly Tool which is freely available online. Its simple interface can guide you through both experimental...with multiple point mutations NEBuilder HiFi can simplify multi-position site-directed mutagenesis by assembling...desired mutations. This same process can be used simultaneously for the generation of one or more mutations...With NEBuilder HiFi DNA assembly, it becomes as simple as mixing fragments together with linearized vector...laborious, requiring multiple cloning steps. You can simplify this process by using NEBuilder which requires...oligo. Generating an sgRNA construct becomes as simple as combining your favorite linearized plasmid with...

...confidence that they aren’t simply doing an experiment wrong. However, even simple procedures like pouring...Reproducibility Project: Cancer Biology have produced similarly low rates of reproducibility. Failed attempts ...someone else's new and incredibly useful technique simply by following the abbreviated methods section found...modified analyses on published data if you need to. Similarly, FigShare also makes it easier to share data through... Figure 1 where many scientists are performing similar experiments. Biology is messy enough that by chance...consolidate published and unpublished experiments on similar subjects in a single, searchable space. Finally... more likely that we’ll find a fluke when we’re simply trying to get our experiments to work, but part...

.... Thank you to our guest blogger Derek Simon! Derek P. Simon received his PhD in Cellular and Molecular...composer. View his blog drsimonsaysscience.org or follow him on Twitter @derekpsimonphd.    References 1. Gutierrez...This post was contributed by guest blogger Derek Simon. The surgeries and standard molecular neuroscience...cemented to the animal’s skull using dental cement (a similar procedure as implanting a guide cannula). A fiber... which can control output of several lasers simultaneously, can be used to set the laser output without... Central PMCID: PMC3769938. 5. Iwai Y, et al. A simple head-mountable LED device for chronic stimulation...

... your lab bench, we need to (this is a little simplified): Find the appropriate glycerol stock within...that everything will go perfectly and our software simply follows a linear progression directing our techs... the most failure prone. In an academic lab, a similar situation could arise if a lab veteran who worked...failure: In our second example we’ve added some simple but critical logic. Some steps fail “hard”, they...the whole procedure. Other steps fail “soft” they simply need to be repeated. We also now have a good interface...contents from the container (of course this is a simple task in the lab, but the software should be mirroring... growth strains, vector backbone, etc.) and you simply don't have enough context to know what they really...

...provide you with some tips on how to automate some simple processes in your own lab. Automating bacterial...up a basic protocol on the Biomek FxP is pretty simple. By clicking a few buttons you can quickly tell...time. It follows all the same steps and uses the similar buffers as would be used for a mini prep, except...to automate these processes, but there are many simple steps you can take to automate processes in your...project in which you need to do a lot of aliquoting. Simply program the pipette with how much total sample ...Cap tubes with a micronic cap sealer. This is a simple table top device that has a roller to go over caps... of pushing them in one by one. It seems very simplistic and it isn’t electronic but it is one of the ...

...protein variants, Cas13, and Cas12a, to develop simple, portable, and inexpensive platforms to reliably...be detected on commercial lateral flow strips, similar to pregnancy tests. With lateral flow strips, the...DNA or RNA of interest within a given sample is simply determined by the number of bands present on the...CRISPR Cas variant that can also cleave dsDNA similarly to Cas9 (Zetsche et al., 2015 ). Cas12a however...DETECTR. How does DETECTR work? DETECTR works similarly to SHERLOCK. Cas12a is targeted to a specific ...DETECTR could accurately distinguish between two similar types of HPV, HPV16 and HPV18, from human cells...Applications of DETECTR in diagnostics DETECTR allows for simple and efficient detection of nucleic acids in a mixed...

...coming weeks, but, in this post, we will outline a simple method for selecting the guide RNA, validating ...CRISPR Design tool supplied by the Zhang lab. This simple website invites you to input the target region ...happy with your selection, ordering the sgRNA is simple and easy. Several companies supply gRNA sequences...you gRNA in vitro first. In vitro screening is a simple and fast method for validating your selected gRNA...). Testing the efficacy of your gRNAs is then a simple matter of transfecting HEK293T cells with your ...the individual gRNAs in their PX330 plasmids (or similar plasmids for gRNA expression), one gRNA at a time...perfect primer sets to use to genotype your pups. A simple PCR with these primers will let you know if there...

...sometimes localize to unintended sites with sequence similarity to the on-target site. When a CRISPR-Cas system...targets by selecting a gRNA with low sequence similarity elsewhere in the genome. There are many predictive...Once you’ve selected a gRNA with low sequence similarity elsewhere in the genome to reduce off-targets...to consider your Cas enzyme. Even if there are similar sequences present in the genome, certain Cas enzymes...Candidate site sequencing Sites with high sequence similarity to the gRNA can be sequenced. These sites can...appropriate method for an activator or repressor Cas. Similarly, Cas ChIP-Seq would also identify all DNA binding...experiment due to their location (e.g. in a gene in a similar pathway, a growth factor, etc.) this control is...