...two loxP sites, which can be located either on the same or on separate pieces of DNA. Both 13bp repeat sequences... sites: Inversion: If the loxP sites are on the same DNA strand and are in opposite orientations, recombination... reversed.   Deletion: If the sites face in the same direction, the sequence between the loxP sites is...sites undergo recombination with other sites of the same type, but are not cross-compatible. Using different...

...don’t use the same overhang twice; (2) avoid palindromes; (3) no overhangs with the same three nucleotides...in a row; (4) no more than two nucleotides in the same position; (5) avoid overhangs with either 0% or ...71% (Pryor et al., 2020). And in 2022, using the same tools, they successfully assembled a 40 kb T7 phage...

...corresponding gene products due to the same region, but not the same exact genes, being improperly silenced.... This is how most cells in the body contain the same DNA but have very different structure and function...present without any DNA changes, and both involve the same region of chromosome 15. They both result from the...

...scientists design and analyze batches of CRISPR samples. We invite you to jump in and try it out, or take... databases, including primer3, biopython, bwa, samtools, bowtie, UCSC genome browser, Ensembl, and others...currently Crispor and Crispresso, that adhere to the same principles listed here. It employs biological best...productivity in the Biohub. We hope it will do the same for you. Send us your questions or comments to gdingle...

...particles that are present in an AAV prep. AAV samples are first DNaseI digested to remove any residual...technology or a TaqMan primer/probe set can be used and samples are quantified by comparing to an AAV standard ...AAV, remember to account for the dilution of the sample for the DNAseI digest. Use good sterile technique.... 2. Choi, V.W., Asokan, A., Haberman, R.A., & Samulski, R.J. (2007). Production of recombinant adeno-...

...between endogenous and exogenous versions of the same target. In the 1980s, Munro and Pelham set out to... ideal.  Unexpected connections Right around the same time that Munro and Pelham were devising their tagging...from this year.  But are we all really using the same antibody developed almost 40 years ago? Well…kinda...distributing monoclonal antibodies derived from the same hybridoma developed by Evans, et al. In other cases...

...fluorescence when a small magnet was passed over the sample (Frank Hayward et al., 2024). The fluorescence ...intensity decreased whenever the magnet approached the sample (by less than 1%) and recovered after the magnet...expressing MagLOV while a magnet is waved under the sample. The imaged area measures approximately 82 mm across...Matsuoka, R., Kimura, S., Miura, T., Ikoma, T., & Kusamoto, T. (2023) Single-Molecule Magnetoluminescence...

...beam into a thin sheet to illuminate a fluorescent sample and captured the reflected light using a different...laser and the reflected light travel through the same objective lens. Using this method, these authors...microns, illuminates only one thin plane of the sample surrounding the focal plane of the detection lens...problem is to illuminate and acquire images of a sample from two opposing sides and merge the images, such...

...laboratory. All procedures are performed using the same three step protocol:  Fragment generation - Any...events, all linear fragments need to get into the same cell. For simple cloning requiring just one or two...vector and insert with separate primer pairs in the same PCR. Add homologous sequences to one of the primer...simply by adding all the necessary primers to the same PCR mix. Pairs of primers will amplify the whole...

...future.  Many thanks to our guest blogger Samantha Young. Samantha Young is a Medical Writer with a PhD in... This post was contributed by guest blogger Samantha Young. The use of CRISPR/Cas9 for gene editing has...technique, you can assess several sgRNA’s at the same time, and choose the most effective ones to increase...

...reintroduction, inversion and re-inversion happen at the same probability. Continuous inversion of the DNA sequence...recombination can only occur between target sites of the same sequence in FLEx vectors, the DNA sequence that ...neuron and its synapse should be visualized at the same time from two different plasmids this approach becomes...connectivity before and after the gene knockout in the same mouse.   Scientists always strive for more....

...thanks to our guest blogger Samuel Mortensen from Northeastern University. Samuel Mortensen is a PhD candidate...This post was contributed by Samuel Mortensen, a PhD candidate at Northeastern University. Working with...19457242. PubMed Central PMCID: PMC2693113. Mortensen, Samuel, et al. "EASI transformation: An efficient transient...

...our metabolic engineering case study of N-acetylglucosamine biosynthesis, we constructed a strain library...rounds of transformation to complete the N-acetylglucosamine strain library construction in 8 days. To ...numbers. (D) The effect of gene dosage on N-acetylglucosamine concentration. Image from the Yang lab. ... a lecturer in the College of Life Sciences that same year. Her research is focused on the escape mechanism...

... are represented as a series of dot plots in the same order. However, in an actual flow cytometry run,...parallel.  Let’s imagine we’re looking for cells in a sample that are negative for marker A and positive for... proxy for nucleic complexity of the cell.  The sample will have been labeled using a panel of markers... dots. In a contour plot, each line contains the same number of events. Thus, contour lines that are close...

...and subsequently leading to a positive signal. The same effect can be achieved for a fluorophore that relies...context of a paired fluorophore protein. Years later, Samie Jaffrey’s laboratory took advantage of their deep... Jaffrey lab asked:  Could an RNA aptamer do the same thing? Using the HBI derivative 3,5-difluoro-4-hydroxybenzylidene...· February 2008. 11. Svensen, Nina and Jaffrey, Samie R. “Fluorescent RNA Aptamers as a Tool to Study ...

...al., 2008) AAV6 (Towne et al., 2010), AAV8, -9 (Masamizu et al., 2011); AAV5 (Aschauer et al., 2013). Anterograde...PMCID: PMC5529245. Choi, V.W., McCarty, D.M., and Samulski, R.J. (2005). AAV Hybrid Serotypes: Improved Vectors...PMC2895694. Hollis, E. R. 2nd, Kadoya, K., Hirsch, M., Samulski, R. J., and Tuszynski, M. H. (2008). Efficient...PMID: 25374356. PubMed Central PMCID: PMC4269290. Masamizu, Y., Okada, T., Kawasaki, K., Ishibashi, H., Yuasa...PubMed Central PMCID: PMC5572074. Salegio, E.A., Samaranch, L., Kells, A.P., Mittermeyer, G., San Sebastian...

... purified from the system, researchers can run samples on SDS-PAGE gels containing Phos-Tag, a small molecule...non-phosphorylated serine at the TAG codon of the same expression vector used for phosphoserine incorporation...incorporation of serine) population of protein using the same expression plasmid. This is a particularly important...express the completely wild type protein from the same plasmid.  Upon verifying that serine phosphorylation...

...seems obvious, it is important to remember that the same is true when designing gRNAs for using CRISPR technology...an alternative to HDR for introducing edits. The same locational constraints are even more exquisite for...concordance between different sequences targeting the same gene, suggesting that off-target effects did not...multiple gRNAs of different sequences give rise to the same phenotype in order to conclude that the phenotype...

...between samples. In addition to a DNA ladder standard, it is also a good idea to run an uncut sample of each...you to either stain your gel after you run your samples or add the stain as the gel is being made (post...containing ligation mixture with insert and backbone. Sample results indicative of successful and unsuccessful...

...campus speakers thoughtfully reminded us to use the same critical lens on our scientific community as we ... nor would you mention my small error and at the same time pass over in silence his very large one” (http...exoplanets into a successful PhD thesis. We use the same atmospheric physics to understand exoplanets that...happy to see so many others come out who feel the same.  I was also incredibly impressed with how well ...