...OZ551 and OZ552 OPEN OPEN gGTCATCTGCcattcGAGGCGGACa REST corepressor 2 OZ553 and OZ554 OPEN OPEN gCTCACCATCtgttggaTGCGATGGAa...

...When cloning by restriction digest and ligation, you use restriction enzymes to cut open a plasmid (backbone... of how to do perform restriction digests.   Overview of the restriction cloning process. Both ...) and the DNA sequence of interest (green, insert) are cut with restriction enzymes to generate compatible...backbone will contain a variety of restriction enzyme cut sites (restriction sites) downstream of the promoter...DNA (insert) that has been cut by compatible restriction enzymes. An enzyme, DNA ligase, then covalently...backbone covalent.  Before beginning the restriction digest and ligation process, you should carefully...these both must have compatible cut sites for restriction enzymes that allow your insert to be placed into...

...methylation may affect your restriction digests. Why methylate? Restriction enoducleases are so ubiquitous...It turns out that restriction enzymes are one half of naturally occurring restriction modification systems... from the restriction modification systems, yet can still affect whether certain restriction enzymes can...associated with any restriction modification systems, their sequences may overlap with restriction sites, inhibiting...Have you ever tried digesting with XbaI or ClaI restriction enzymes and gotten unusual or unexpected results...sequences to prevent them from being degraded by restriction endonucleases. A given prokaryote typically has... has genes encoding one or a few restriction modification systems containing methyltransferases that add...

...allows for the isolation of a genomic region of interest targeted by a gRNA. CRISPR-mediated purification...3xFLAG-dCas9) and a gRNA targeting their genomic locus of interest, the promoter region of the transcription factor... region alleviates these issues as a region of interest can be targeted using a gRNA, which can easily...system can be used to isolate genomic regions of interest with dCas9 and a targeting gRNA. One such example...

...transcription during vertebrate embryogenesis. My interests, though, have proven unruly. Instead of settling...terms of both types and topics of work. A younger interest in being a writer kept chirping up. The evolutionary...In my role as an editor at Science, my various interests come together as useful and productive, forming...forming a different constellation of interests and skills than what fits for applied research. I focus in ...Science communication opportunities If you are interested in the science communications world, you might...more planets to choose from. Put your skills and interests into your backpack, and step out there into the...

...Adult Dermal Fibroblasts Neurons Lentiviral Human REST suppression mediates neural conversion of adult ...

...plasmids. Here, we'll cover restriction digest analysis. Diagnostic restriction digest Diagnostic digests...Diagnostic restriction digests are comprised of 2 separate steps: 1) incubating your DNA with restriction enzymes...quick overview of how to analyze a restriction digest:     Restriction digest tips and tricks: The following...Congratulations, you have a plasmid expressing your gene of interest (YGOI) and are ready to dive into your functional...organization of different features within the plasmid. Restriction analysis can also be used successfully even if...relative sizes of the resulting DNA fragments. Restriction digests are commonly used to confirm the presence...backbone. To do this, you'll use enzymes with restriction sites that flank the insert. You will need to...

...you’re on a break from expressing some genes of interest).   Blugene is so excited they can’t stop...

...Plasmid Cloning by Restriction Enzyme Digest (aka Subcloning) You may also like... Restriction Digest of Plasmid...identify which restriction sites are present in a given sequence. When selecting restriction enzymes, you...desired restriction sites to flank your insert : You can use PCR Based Cloning and add restriction sites...need to avoid restriction enzymes that cut within your insert. Adding desired restriction sites to your... same restriction enzyme buffer (see New England Biolabs for more information about restriction enzyme...Information about plasmid cloning by restriction enzyme digest (subcloning), including design and experimental... another as long as it is already bounded by restriction sites that are also present in the same orientation...

... Protocols Restriction Digest of Plasmid DNA Restriction Digest...a list of many commonly used restriction enzymes, visit NEB . Restriction enzyme digestion is commonly...of your plasmid of interest (see below for recommend amounts) Appropriate restriction enzyme (see manufacturer's...Procedure Select restriction enzymes to digest your plasmid. *Pro-Tip* To determine which restriction enzymes ...typical restriction digestion reaction could look like this: 1 µg DNA 1 µL of each Restriction Enzyme ...cleavage at certain restriction sites. See NEB's table of methylation sensitive restriction sites . Sometimes...perform a restriction digest of plasmid DNA Protocols...

...pGP-AAV-syn-FLEX-jGCaMP7b-WPRE) exhibits the brightest resting fluorescence of the GCaMP7 family of calcium sensors...exhibits high contrast between peak fluorescence and resting fluorescence. It is useful for activity imaging...exhibits high contrast between peak fluorescence and resting fluorescence.  It is useful for activity imaging...particular cell types. For example, the mDlx enhancer restricts expression to GABAergic interneurons. These vectors...

...peak fluorescence and resting fluorescence (achieved through lowering sensor resting fluorescence, while...responding variant. jGCaMP7b exhibits the brightest resting fluorescence and can be used for imaging small ...

...access to the antibody sequences, in line with the rest of Addgene’s recombinant antibody collection, as...

...performer if they are enough of a jerk to make the rest of the team miserable.   A good manager keeps...dissatisfaction, the work can get done, and get done well. The rest of the blogs in this series will address more specific...

...out our referral form and we’ll take care of the rest. Know of a useful plasmid that’s not in the collection...

... China was affected by COVID-19 earlier than the rest of the world, from mid-January to mid-March.  To... of all of our plasmid orders, we will focus the rest of our discussion on these three regions. Each of...indefinitely due to COVID-19. In our lab, we were interested in examining the effect of COVID-19 on worldwide...graduate student at Harvard University. He is interested in using selection, evolution, and machine learning...

...education) by naming the vectors after him.  The rest, as they say, is history: pBabe vectors have been...as a technician) of Bob Weinberg’s lab we were interested in expanding on the concept of oncogene cooperation...

...issued by Google in October entitled “Rise of the Rest: The Growing Impact of Non-Elite Journals” found...for a moment, it’s clear that articles in less-prestigious journals have been gaining ground. A study issued...proportion of highly cited articles appearing in lower-prestige journals has risen steadily from 1995 to 2013.... in citations for older publications. (As an interesting aside, Nature recently reported that the most...

...conditions: The sequence is unique compared to the rest of the genome. The target is present immediately...sequence is for the genomic target compared to the rest of the genome. Ideally, a gRNA targeting sequence...nucleotide edits that introduce a novel restriction site. The region of interest is PCR amplified, digested with...region of interest can be PCR amplified using primers that (A) flank the region of interest (deletions...sequence must be delivered into the cell type of interest with the gRNA(s) and Cas9 or Cas9 nickase. The...identify are actually important for your phenotype of interest. You can individually test the gRNAs identified...to ensure that they reproduce the phenotype of interest. Find more information on factors to consider ...

...compared to untagged GFP overexpression, while the rest of the fusions had some negative effect on these...help you determine localization of a protein of interest, purify it, or determine its expression level ... with CRISPR Fluorescently Tag Your Protein of Interest ...