We narrowed to 530 results for: Erf
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TypeBlog PostPublishedJan. 28, 2014, 3:10 p.m....We perform over 100 transformations each week at Addgene, so we need to be cost and time efficient. We...aliquoted and stored at -80oC until we are ready to perform transformations. A typical 200mL bacterial culture... of the large number of transformations being performed at Addgene, this adds up to a significant time...
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CRISPR/Cas9 FAQs Answered!
TypeBlog PostPublishedMarch 13, 2014, 4:08 p.m....isolate single-cell colonies, grow them up, then perform individual genotyping (using Sanger sequencing,.... You could then confirm this purification by performing a genotyping assay (such as Sanger sequencing... 500ng of 293FT genomic DNA as template after performing gDNA purification and have tried conditions with...recommends using the Epicentre QuickExtract solution to perform this step to extract genomic DNA from cells. The... -
Zebrafish as a Model for Behavior: Swimming into the Optogenetic Spotlight
TypeBlog PostPublishedJuly 16, 2019, 12:59 p.m....react by performing a distinct C-bend to escape possible danger (see figure). Larvae also perform spontaneous...effects on behavior. The system is an especially powerful tool for in vivo behavioral studies because it... -
Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process
TypeBlog PostPublishedFeb. 1, 2018, 3:07 p.m....how we use next generation sequencing results to perform quality control on deposited plasmids. We’ve also...improved feature detection with an easy to use interface. We regularly use Snapgene for our quality control...region is present in the plasmid, we might need to perform a restriction digest to confirm the size of the... -
SunTag and Fluorescent Imaging
TypeBlog PostPublishedMarch 28, 2017, 2:30 p.m....various conditions. However, this system is far from perfect. For example, If your fusion protein is expressed...GFP aggregation, which they reduced by using superfolder GFP (sfGFP) with the small solubility tag GB1...What can you do with SunTag? Tanenbaum et al. performed a variety of different experiments in their paper... -
FastCloning: A Newer, Simpler Cloning Technique
TypeBlog PostPublishedNov. 1, 2016, 2:30 p.m.... While restriction cloning is still routinely performed by many labs, a variety of other cloning techniques...transformation. After transforming cells, you should perform colony PCR with vector specific primers to confirm...Resources on the Addgene Blog Plasmid Cloning by PCR Perform Site Directed Mutagenesis by PCR Catch up on Your... -
Antibodies 101: Epitope Availability
TypeBlog PostPublishedJan. 11, 2022, 2 p.m...., changing locations, or even just as part of performing a specific function. A transmembrane protein ... validated for the type of assay that you are performing. If they’re not available, the next best thing...denatured protein samples. If, however, you are performing an immunohistochemistry assay, you may want to... -
Identifying Sequence Elements with SnapGene's Feature Database
TypeBlog PostPublishedDec. 21, 2017, 2:06 p.m....painless as possible. Instead of crowding the interface with every possible option, we place the most ... identifying a feature even when the match is imperfect. SnapGene's feature database The source of common...detection system was enhanced to allow searches for a perfect protein sequence match even when the DNA sequence... -
Multifunctional Neural Fibers: Modulation from Gut to Brain
TypeBlog PostPublishedAug. 31, 2023, 1 p.m....channel, fused to mCherry to the brain tissue. They performed optical stimulation using NeuroStack and the fibers...to me. What’s on the fiber horizon? The brain interfaces with the entire body, so the obvious next applications...Flexible fiber-based optoelectronics for neural interfaces. Chemical Society Reviews, 48(6), 1826–1852. ... -
What Do I Do Now? Academic v. Non-Academic Career Decisions
TypeBlog PostPublishedMarch 9, 2017, 3:40 p.m....of today’s postdoctoral training can directly interfere with a seamless jump into industry.” In his article...opportunities—projects end and layoffs occur, but good performers move around easily Table 1: Taking it a Step... career with advancement in one company, good performers in for the long haul Limited movement in one... -
Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?
TypeBlog PostPublishedJune 9, 2016, 2:30 p.m....Argonaute proteins play an important role in RNA interference, binding RNA guides to cleave foreign RNA. Argonaute...choosing a genomic target. NgAgo also appears to outperform Cas9 in GC-rich regions. DNA guides NgAgo uses... 2. Swarts, Daan C., et al. “DNA-guided DNA interference by a prokaryotic Argonaute.” Nature. 507(7491... -
Plasmids 101: NGS Quality Control for Pooled Libraries
TypeBlog PostPublishedOct. 26, 2017, 1:59 p.m....Addgene uses electrocompetent Stbl4 cells when performing amplifications, and we use electroporation to... distribution, with an AUC of 0.5 representing perfect uniformity (see Figure 3A). Addgene finds that ...pooled plasmid libraries. Pooled libraries can be a powerful research tool, but their proper use requires careful... -
Using AAV for Neuronal Tracing
TypeBlog PostPublishedAug. 9, 2018, 1:04 p.m....of AAV could facilitate the development of more powerful AAV serotypes, and perhaps lead to controlled ...neuronal networks, improved AAV vectors could also be powerful tools for the field of gene therapy if they can..., Phillips, J.W., Hohmann, J.G., Wohnoutka, P., Gerfen, C.R., Koch, C., Bernard, A., Dang, C., Jones, ...Michael, S., Kuleshova, E., Ojala, D., Huang, C.C., Gerfen, C.R., Schiller, J., Dudman, J.T., Hantman, A.W... -
15 Hot Plasmids from 2017
TypeBlog PostPublishedJan. 9, 2018, 3:02 p.m....separable from that of EGFP. This makes mCyRFP1 a powerful tool for use as a donor in FRET (fluorescence ...residues on the outer surface to break dimerization interfaces (affecting many of the RFPs developed from natural...All three mScarlet variants have shown great performance in cellular functional imaging and in protein...of essential genes in vivo. Utilizing CRISPR interference (CRISPRi) technology, the arrayed knockdown ... -
Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering
TypeBlog PostPublishedMay 3, 2018, 1:48 p.m....have taken the CRISPR world by storm. Like RNA interference, these enzymes can knock down RNA without altering...nuclease domains characteristic of the Cas13 superfamily. Yan et al. proceeded to study orthologs from...multiple RNA regulating methods: small hairpin RNA interference, dCas9-mediated transcriptional inhibition (... -
Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)
TypeBlog PostPublishedSept. 26, 2024, 1:15 p.m....but it doesn’t really matter if those overhangs perfectly match up. They just need enough complementary ... To overcome this limitation, one option is to perform a hierarchical assembly, assembling fragments in...Choosing a Molecular Cloning Technique Choosing Your Perfect Plasmid Backbone ... -
Hot Plasmids - February 2022
TypeBlog PostPublishedFeb. 22, 2022, 2:15 p.m....-based method for screens. Cas12a’s ability to perform multiplex editing within a single expression cassette...used to create a genome-wide library capable of performing similarly to Cas9 screens in a compacted library... via a nuclease-active CasMINI. This small yet powerful system opens up new possibilities in both cell... -
Synthesized by Ginkgo Bioworks, Shared by Addgene: SARS-CoV-2 Plasmids for Many Expression Systems
TypeBlog PostPublishedMay 19, 2020, 1:15 p.m....lab team manually transforms, innoculates, and performs DNA minipreps for small batches of samples. As...standard pipeline. For the first ~50 plasmids we perform our standard full plasmid sequencing by NGS. Addgene...and make them available to scientists without performing the full suite of time consuming, costly, and... -
Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools
TypeBlog PostPublishedDec. 6, 2018, 1:34 p.m....Listen to the C. elegans RNAi podcast segment RNA interference (RNAi) is extensively used in C. elegans research...endogenous RNAi pathways. These mutations however can interfere with other genetic pathways making it difficult...experiments. Potential experiments that one might perform using the purified domains include protein stability... -
A Practical Approach to Choosing the B(right)est Fluorescent Protein
TypeBlog PostPublishedJune 1, 2017, 2:30 p.m....couple of promising variants to check how they perform under your experimental conditions. This is time...combinations of biology and equipment. Because the performance of a fluorescent protein strongly depends on ...endogenous tagged genes) can be used to verify performance under different imaging conditions (e.g. different...