...targeting constructs. In the example shown in Figure 1, large sequences corresponding to exons 1 and 3 of the ... the cells that have randomly integrated all or large portions of the plasmid. Deliver your construct ...CRISPR is very good at making knockouts, knocking in large sections of DNA can be more difficult. Addgene depositors...

...strains, including HB101 and derivatives, release large amounts of carbohydrates that can inhibit the cell...levels of protein, not plasmid DNA. If you need large amounts of DNA from a plasmid in a protein expression...gas exchange in the overnight culture. Generally, large cultures in flasks may be shaken around 220 RPM,...coli strains. Increase the volume Starting with a larger culture volume can improve plasmid yield. Our lab...

...change in genotype. Epigenetic modifications play a large role in turning genes ‘off’ and ‘on’, signaling ...Epigenetic changes have been discovered to play a large role in a variety of diseases, ranging from cancer...changes in their DNA methylation patterns, either large increases or decreases. Hypermethylation of CpG ...

...is often the case with large genes. This is due to inefficient packaging of large inserts; a study by Kumar...start. If you don’t anticipate that you will need large volumes of lentivirus routinely, you may opt for... guides) while a single 72h harvest is best for larger genes such as Cas9. See figure 1 for an example...dishes or increasing the vessel size. Opting for a larger vessel, such as a 3 or 5-layer flask will require...calculating the ratio of the surface area of the larger vessel to the 10 cm2 dish and adjusting the cell...

... to take next steps and follow up especially in large companies. It is fine to address the cover letter...might be able to help you. If the organization is large, you will need to be more directed and target someone...

...United States, I look forward to gathering around a large table for a meal with my family. I am thankful for...things, and one is that I get to gather around a large table to eat with my Addgene family every day. Do...

...removing the full ncRNA locus (Ho et al. 2012). For large-scale screening, sgRNA libraries are useful for ... same time (Liu et al. 2018). However, removing large lncRNA fragments with multiple gRNAs poses the risk...Mutations of splice sites. (C) Removal of an exon or a large genomic fragment. (D) Knock-in: insertion of a synthetic...

...researchers and ready access to stably archived large-scale datasets by computational biologists. ...genome-wide datasets, which are often obscured in large supplementary data files, BioGRID ORCS makes it ...

...paper and then another figure in another paper. A large fraction of readers cover the same ground again ...allows you to make meaningful collections from a large section of open access life sciences data. You can...

...his team created with the research community at large, "[Fluorescent proteins are] a whole new way of ...scientists, and the FSU team who have made this large, important plasmid deposit possible.   ...

... SPH mice CRISPRa has largely been limited to in vitro settings due to the large size of Cas9 activators...activators, which largely precludes delivery by adeno-associated vectors (AAV). This issue was recently circumvented...delivered to the CNS by lentivirus, which has a larger packaging capacity than AAV. Such an approach was...

...Additionally, CAPTURE 3C-based sequencing requires a large number of cells (~5x107). Together, these two requirements...interactions identified with the multiplexed CAPTURE 2.0 largely replicated interactions previously identified with...chromatin interaction networks since they are a large cluster of enhancers. CAPTURE 2.0’s multiplexing...

...into the genome, and its large DNA packaging capacity is well suited for larger transgenes. Unfortunately...defects in the gene MYO7a, at ~7 kb this gene is too large to deliver via AAV.  Dual vectors strategies are... to regulate and optimize transgene levels, and large scale manufacturing of viral vectors to support ...cargo - 4.8 kb at most - which means that a gene larger than 3 kb cannot be delivered using this vector...

... in the lab. Advantages of lentivirus include a large genetic capacity and the ability to transduce both...the clinic Since lentiviral vectors can deliver a large amount of DNA (~8 kb) with a relatively low immune...lentiviral vectors for applications that require the large genetic capacity of a lentiviral vector but for ...Dorazio R, Mulder GJ, Polack A, Bank A, Soulier J, Larghero J, Kabbara N, Dalle B, Gourmel B, Socie G, Chrétien...

...III-E, and then used this information to identify a large variety of related bacteria with similar Cas systems...fused integrase would then integrate into with a large DNA insert – or Programmable Addition via Site-specific...programmed for new genes. In comparison with other large insert techniques like HITI or HDR, which require... and adenovirus templates. The system, which is larger than Cas9, can also be used with mRNA and a synthetic...

...tools for efficient and programmable insertions of large DNA cargos into bacterial genomes. This new capability...CRISPR-transposons excel at programmable insertions of even very large genetic cargo, at one or multiple desired genomic...systems have already been demonstrated, ranging from large genomic insertions and/or deletions, to multiplexed...

...detection; however, those are just one (admittedly large) class of common fusion protein tags. Biochemists...; has inherent enterokinase cleavage site GST Large Protein 26 Purification and Stability Good for purification..., flanked by hexahistidine motifs (6xHis) MBP Large Protein  40 Solubility and Purification Can improve...

...observed to occur more frequently in plasmids with large inserts (>7 kb)(Berg et al., 1989), direct repeats...reads (<300 bp) which cannot distinguish between large repeat sequences within a plasmid, it cannot be ...sequencing. These longer reads are able to detect large repeat sequences within a plasmid and, therefore...DE. pBR322-derived multicopy plasmids harboring large inserts are often dimers in Escherichia coli K-12...just as well as monomers. However, due to their larger size, multimers may exhibit reduced transformation... & Sherratt, 1984; Summers et al., 1993). These larger plasmids are also maintained at lower copy numbers...monomeric version. However, undigested multimers are larger than monomers and will therefore run higher on ...

... that cellular mismatch repair contributed to a large number of unintended prime editing outcomes, including...combine prime editing with a recombinase to install large sequences (up to 10 kb) into the genome (Pandey ...deletion, replacement, integration and inversion of large DNA sequences with twin prime editing. Nature Biotechnology... (2024). Efficient site-specific integration of large genes in mammalian cells via continuously evolved...J. S. (2023). Drag-and-drop genome insertion of large sequences without double-strand DNA cleavage using...

...overheating remains an issue, which can happen with large proteins, try lowering the voltage and using a longer...you know you loaded a good amount of sample of a large protein, you may be under-transferring.  Membrane...good indication that the p membrane pores are too large.  Transfer method   Wet transfers are more efficient...efficient, so if you’re struggling to transfer a large protein, consider using a wet transfer with a long... lot of antibodies. If you’re planning to use a large amount of a specific antibody, plan ahead and request...harsher stripping buffer Too little signal Large proteins: Incomplete transfer Check transfer with...proteins going completely through the membrane or larger proteins not transferring completely onto the membrane...