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TypeProtocol...bottom. Add 5 mL of Pierce IgG Elution Buffer to the capped Gravitrap column and incubate for 10 min at RT...subsequent centrifugation step Pro-Tip Resin will appear compacted after centrifugation. Discard the flow...column in a new 50 mL LoBind collection tube. Slowly apply the sample to the center of the compact resin bed...exchange and concentration with an Amicon column Apply 15 mL of PBS to the reservoir of a Amicon Ultra-...
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Neuronal labeling with Spaghetti Monster
TypeBlog Post...fluorescent proteins (FP) have also been important in mapping, monitoring, and manipulating neuronal circuits...combining advantages of FPs and epitope tags, smFPs appear to be very effective probes for finely visualising... -
Analyzing CRISPR Editing Results with ICE from Synthego
TypeBlog Post...workflow - analyzing the data - is often under-appreciated or over-looked. Synthego has developed a new...analysis Once the analysis is complete, a new screen appears with a graphical representation of the results ... -
Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor
TypeBlog Post...transduced by PHP.B. The Deverman Lab took a broad approach and searched for genetic difference between the...phenotype in mice. They took a classic genetics approach and interbred C57BL/6J and BALB/cJ mice to map... -
What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program
TypeBlog Post... providing support "for development of tools, approaches, and infrastructure necessary for direct tests...recent). They have diverse traits with good genetic mapping morphology and interesting behaviors. There is ... -
How to Write a Scientific Review Article
TypeBlog Post...papers and then feel ready to write. What actually happened was that each paper taught me a few things and...to read every single paper. I also learned to appreciate the discussion sections of the primary literature... -
Plasmids 101: Gateway Cloning
TypeBlog Post...sequences into multiple vector systems. With the appropriate entry and destination vectors, one can use Gateway...elements to express the gene of interest in the appropriate system Recombines with the entry clone to generate... -
Hot Plasmids - August 2020
TypeBlog Post... have created a yeast-based directed evolution approach with some notable improvements. Directed evolution...specific guide RNA (gRNA) sequences. But these approaches rely on continual expression of both dCas9 and... -
Antibodies 101: Reading a Flow Plot
TypeBlog Post...similar approach would be good for your work. Hopefully this blog helps you with that…and happy flowing... -
Viral Production at Addgene
TypeBlog Post...particles can be identified after negative staining and appear darker than full vector particles. ...replaced with a new vial of early passage cells. Approximately two weeks post-thaw, cell culture supernatant... -
How to Be an Excellent Trainee
TypeBlog Post...easiest to train individuals who are polite, appreciative, and clear about their needs. Remember, your...trainees one day when you are the senior personnel! Happy training! Resources Resources on the Addgene blog... -
CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?
TypeBlog Post...Motif (PAM), the sequence NGG. If these conditions apply to your experimental target, SpCas9 and its enhanced...specificity over a single SpCas9. The drawback of this approach is that two sgRNAs targeting opposite strands ... -
pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs
TypeBlog Post...possibilities for the clinical and therapeutic applications of iPSCs. The generation of human iPSCs (hiPSCs...., Recent technological updates and clinical applications of induced pluripotent stem cells. Korean J ... -
Antibiotic Resistance: An Old Solution but a New Problem
TypeBlog Post...from random mutations in the genetic sequence that happen to be beneficial, and persist as a result. Why ...are susceptible to the drug, while the few that happen to possess resistance genes survive and multiply... -
CRISPR Activation: A Practical Guide
TypeBlog Post...suffer from an inability to easily deliver the appropriate isoform(s) of a given gene, as, many times, the...engineering parameters via a library-on-library approach. Nat Methods 12:823–826 . https://doi.org/10.1038... -
Plasmids 101: Control Plasmids
TypeBlog Post...RNAi or CRISPR, non-targeting controls may be appropriate as they allow you to assess the specificity of...us now revisit our experiment. In Figure 1, it appeared as though the shRNA did not knock down expression... -
Plasmids 101: E. coli Strains for Protein Expression
TypeBlog Post...plasmid-- make sure to maintain this by growing on appropriate media. Note: Purifying your expression plasmid...to make sure your cells have first reached an appropriate density. Cells in the exponential growth phase... -
Fluorescent Proteins 101: When GFP lets you down
TypeBlog Post... proteins.Therefore, GFP is not suitable for applications that require anaerobic conditions. In experiments...protein module that can report on location. This approach enables the study of proteins in cells, but not... -
Deep Mutational Scanning with One Pot Saturation Mutagenesis
TypeBlog Post...that expresses all desired variants of a protein. Applying a selective pressure winnows the pool down to ...technique described in Wrenbeck et al is a PCR-based approach for generating a customizable comprehensive mutagenesis... -
Five Popular Model Organisms, Part 2
TypeBlog Post...Escherichia coli as a Model Organism and Its Application in Biotechnology. In Escherichia coli - Recent...Physiology, Pathogenesis and Biotechnological Applications. InTech. https://doi.org/10.5772/67306 Koornneef...