We narrowed to 320 results for: POR C
-
TypeBlog PostPublishedDec. 7, 2021, 2:15 p.m.... epitope such as c-Myc, GFP or V5 can be used to tag the protein of interest on the C- or N-terminus. ...Their porous surface provides a large area for binding, but antibodies conjugated within the pores are ...used for IP. These proteins bind only to the Fc portion of the immunoglobulin, leaving the antigen binding...or a magnet (magnetic beads), providing a solid support while unbound proteins are washed away. While ...to be adjusted depending on the sample. It is important to remember that sample lysates will also contain...
-
Fluorescent Proteins 101: Fluorescent Protein Timers
TypeBlog PostPublishedMay 4, 2017, 2:30 p.m....predictable kinetics over 14 hours of expression in C. elegans embryos. However, this initial timer was ...timer could be used in vitro, in mammalian cells, in C. elegans, and in Xenopus embryos. Of course, timers...have been around since 2000 when Terskikh et al. reported the production of dsRed E5. This timer predictably... your experimental setup less cumbersome. 3. Portability across multiple systems Terskikh et al. showed...fluorescent timers that change color from blue to red report on cellular trafficking." Nature chemical biology...Chronology of islet differentiation revealed by temporal cell labeling." Diabetes 58.8 (2009): 1863-1868... -
Droplet Digital PCR for AAV Quantitation
TypeBlog PostPublishedJan. 16, 2020, 2:18 p.m....calculate the viral titer: GC/mL = {[(R*C)(1000/V)]*D} R = Reaction volume C = Copies/uL V = Volume of virus ...by ddPCR begins with diluting the virus. It is important to note that the dynamic range of the ddPCR is...PCR plate. Maintaining a high droplet count is important for calculating the concentration of your sample...separation (Witte et al., 2016). This is particularly important if your amplicon is longer than 150 base pairs... -
CRISPR 101: Targeting RNA with Cas13a (C2c2)
TypeBlog PostPublishedSept. 21, 2017, 2:07 p.m....overhangs Cas13a (C2c2) 2x HEPN crRNA RNA 3' A, U, or C (not required by all orthologs) Specific RNA cleavage...spacer sequence and consists of a single A, U, or C base pair. In bacteria, once Cas13a has recognized...of interest and an inactivated fluorescent RNA reporter. If the target sequence is present in the pool...activity of Cas13a becomes activated and the RNA reporter will be cleaved resulting in activation of the...in the field and as always, we appreciate the opportunity to share these invaluable reagents with the scientific... -
Production of Virus in Insect Versus Mammalian Cells
TypeBlog PostPublishedNov. 5, 2024, 8:32 p.m....https://doi.org/10.1096/fj.01-0995fje. Tan, E., Chin, C. S. H., Lim, Z. F. S., & Ng, S. K. (2021). HEK293 ...doi.org/10.3389/fbioe.2021.796991. Urabe, M., Ding, C., & Kotin, R.M. (2002). Insect cells as a factory ...of production, saving about a week of work. An important note is that this is only possible because baculoviruses... will come down to which differences are most important to you — virus type, cost, time, or reagents and... -
Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity
TypeBlog PostPublishedOct. 11, 2018, 12:30 p.m....construct. Reactions are often cycled between 37°C and 16°C to provide optimal reaction temps for both the... Golden Gate assemblies feature an inverse proportionality between the complexity of the assembly (number...The future of assembly DNA assembly methods are important tools for many areas of science, and researchers... -
CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics
TypeBlog PostPublishedAug. 9, 2016, 2:30 p.m....our example later, histones) ChiRP-seq RNA Hi-C DNA (genome-to-genome interactions) DNase-seq DNA...commonly think of DNA as a linear polymer of A,T,G, and C nucleotides arranged in a double helix. People are...genome. But not all of those 13,000 regions are important for maintaining ESCs. So which of these 13,000...next-generation sequencing to measure the relative proportion of guide RNAs (gRNAs) across the population. ... -
Plasmids 101: Protein tags
TypeBlog PostPublishedDec. 11, 2014, 4:26 p.m....tag to it. The FLAG, hemaglutinin antigen (HA), and c-myc tags have been the workhorses of the affinity ... tags, a His tag can be fused to either the N- or C-terminus of a protein. Unlike other epitope tags –... Protein tags are usually smallish peptides incorporated into a translated protein. As depicted in the... are available, but scientists would need to incorporate their recognition sites into their protein tag... -
Plasmids 101: Gateway Cloning
TypeBlog PostPublishedJan. 12, 2017, 3:30 p.m....into an attL-containing entry vector. Method C: Restriction cloning of a restriction enzyme fragment...attL-containing entry vector. Figure 5: Method C to create an entry clone: Restriction cloning the ...coding sequence. If you choose this strategy, it’s important to include the proper protein expression elements...Clone When making the expression clone, it is important to choose the destination vector that best fits... -
Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs
TypeBlog PostPublishedOct. 27, 2015, 3 p.m....create an N- or C-terminal fusion based on your experimental needs. Equally important, before cloning ... retrieval sequence only functions at the extreme C-terminus [3]. You must therefore determine your cloning...potentially affect function [5]. We and others have reported that FP cysteine residues form inappropriate disulfide...