We narrowed to 636 results for: erf
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TypeBlog Post...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 (...
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A Practical Approach to Choosing the B(right)est Fluorescent Protein
TypeBlog Post...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... -
Screening for Successful Genome Editing with Digital PCR
TypeBlog Post...target DNA. Conventional thermal cycling is then performed to amplify the target using PCR. Instead of tracking...mutation status. The drop-off probe is designed to perfectly complement the wild type sequence and is not capable...design such assays, in addition to testing their performance (2). Remember that although digital PCR is a ... -
Hot Plasmids - February 2022
TypeBlog Post...-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 Post...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 Post...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... -
To Each HIS Own
TypeBlog Post...step to remove the tag in order to prevent it interfering with protein function. In contrast, the polyhistidine... it is, and the greater the risk that it will interfere with protein function. Additional Benefits of...this one developed by Dr. James Trimmer’s lab, to perform assays that they otherwise would not be able to... -
All in a Twist: dsRNA
TypeBlog Post...stranded? Well, that’s just for DNA – right? RNA performs almost all of its biological functions in our ...sense dsRNA and respond accordingly. While not perfect, these sensing systems work to distinguish between...blog CRISPR 101: RNA Editing with Cas13 RNA Interference in Plant Biology ... -
Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics
TypeBlog Post... forces scientists to use invasive measures to perform in vivo optogenetic experiments. Intracranial injections...correlated with the model’s predictions. Three top performers, named ChRger1, ChRger2, and ChRger3, were then...minimally-invasive optogenetics While ChRgers performed well in a tissue culture dish, that doesn’t guarantee... -
27 Hot Plasmids from 2016
TypeBlog Post... from Addgene’s repository that can be used to perform screens in human cells. General information regarding...Liu at Oxford University, we are now providing a powerful CRISPR-based screening source for the Drosophila...resources for fly labs who wish to harness this powerful genome engineering tool. Bassett, et al. J Genet...locus causes that locus to be methylated. This powerful tool has been used to direct methylation to and... that high-throughput tagging projects can be performed. Schwartz, et al. Genetics. 2016. PubMed PMID:... -
Plasmids 101: Modular Cloning
TypeBlog Post...cloning strategy designed by synthetic biologists to perform large scale, one-tube cloning reactions. In this...courtesy of Rob Hurt. What makes MoClo systems so powerful is their modularity — many parts can be assembled...used to assemble multigene constructs capable of performing production and subsequent necessary modification... -
Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them
TypeBlog Post...Andrews et al., 2019). Finally, scientists can perform the entire process of antibody selection and maturation...recombinant antibodies. The selection process is often performed via phage display, yeast display, ribosome display...they are using and any validation experiments performed will be useful in perpetuity. Second, plasmids... -
The Materials Science of Optogenetics Experiments
TypeBlog Post...specific to lasers or optogenetics so cheaper, less powerful alternatives to the Master-9 that only control...published first. Because behavioral optogenetics is performed in freely moving animals, the fiber optic patch...into some of the important considerations when performing optogenetics behavioral experiments. Thank you... -
PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs
TypeBlog Post...thousands of designs in parallel. To do this, we performed high-throughput ‘self-targeting’ screens with ...PRIDICT2.0 score. For the best prime editing performance, we suggest that users select some of the best...learning–based model with high generalization performance. Science Advances, 5(11), eaax9249. https://doi.org... -
Viral Vectors 101: Voltage Indicators
TypeBlog Post... here!). The only issue with these GEVIs? They perform poorly in two-photon conditions, which are necessary...the fluor via FRET. This system is particularly powerful for in vivo imaging especially since JFs can cross...type of interest. Viral pseudotyping is also a powerful tool that swaps capsid proteins from different... -
Transferable Skills Guide: Managing a Team
TypeBlog Post... have a much better chance of creating a high-performing team - which is every manager’s dream. Learn ...often associate feedback with end-of-the-year performance review, but feedback should be given throughout...the year and in a timely manner. If someone’s performance or behavior is inadequate, you need to address... -
Plasmids 101: Knockout/Knock-In Plasmids
TypeBlog Post...One of the most powerful strategies to investigate a gene's function is to inactivate, or "knockout",...arms. When we select cells later, we will first perform a positive selection for the antibiotic resistance... knockout and new CRISPR tools continue to be perfected, the production of cells and mouse knockout lines... -
Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR
TypeBlog Post... capture of chromatin interactions The Xu lab performed a proof-of-concept experiment to show that CAPTURE...original CAPTURE method. Both CAPTURE methods outperformed other 3D chromatin methods such as ChIA-PET ...Wang, K. (2018). 3C and 3C-based techniques: the powerful tools for spatial genome organization deciphering... -
Plasmids 101: 5 factors to help you choose the right cloning method
TypeBlog Post...consider Gibson cloning. In this technique, you first perform a round of PCR to generate DNA fragments with 15...function while 5’ adenosine overhangs left by Taq perfectly anneal with the topoisomerase I generated overhangs...available cloning kits that one could choose from to perform these experiments. In most cases, there will be... -
When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells
TypeBlog Post...monomeric FPs. EGFP and other GFP family members (superfolder, Emerald, CFP, YFP, BFP, Cerulean, Turquoise,...dimerization. Importantly, all of these assays are performed in vitro. Assays to detect FP dimerization in ... 1 and 2 depend on protein concentrations. An imperfect solution is to restrict imaging analysis to cells...