...selected the optimal promoter for your gene of interest, painstakingly cloned into the perfect empty backbone...DNA, important when trying to cut with certain restriction enzymes (ex: ClaI or XbaI) dcm DNA cytosine...DNA, important when trying to cut with certain restriction enzymes that are methylation sensitive. dnaJ...plasmid maintenance, blue/white screening, partly restriction-deficient; good strain for cloning repetitive...methylated, allows for methylation sensitive restriction enzymes to cut the plasmid after preparation....

...protein of interest, and a negative control sample that does not express the protein of interest....detection using antibodies specific to the protein of interest. This protocol describes the basic steps for lysing... Precast SDS-PAGE gel SDS-PAGE running buffer Prestained protein ladder Ethanol iBlot 2 PVDF Mini Stack...depending on the cellular location of the protein of interest. RIPA buffer is suitable for most proteins but...the samples into the gel. Load 5–10 µL of the prestained protein ladder. Place the lid on the tank and...sample type and cellular location of the protein of interest. You may need to try a variety of lysis buffers...

...cloning sites for easy cloning of your genes of interest. In order to improve the study of specific neuronal...new AAV approach using regulatory elements that restrict expression to this cell type. To this end, they... ATP1A1 in addition to a researcher’s locus of interest. NHEJ- or HDR-induced mutations in ATP1A1 can ...locus increases the probability that the locus of interest has also been edited! Thus, ouabain selection ...the size of DNA fragments. Depending on which restriction enzyme is used to cut the plasmids encoding the...are shared through Addgene without licensing restrictions. Use these instructions to prepare the PSU ...

...pediatric kidney cancer. This was particularly interesting as I had just completed my thesis on the study... taking on a similar position in your field of interest before you dive straight into a full-time position...for bacterial contamination. It was a really interesting job at first, everything was very new, and everything...to help cover the costs. My MSc courses were interesting and luckily genetics is a sector which has been...the company as well and use my other skills and interests. I was so glad when my future supervisors were...

...developed the plasmids, pPSU1 and pPSU2. When restriction digested with PstI or EcoRV, these plasmids generate...significantly cheaper than commercial ladders. Are you interested in making your own DNA ladder? You can find the...available from Addgene. Learn How to Analyze a Restriction Digest References 1. Henrici, R. C., Pecen, ... Miniprep Protocol Learn How to Analyze Your Restriction Digests Make Your Agarose Gel Other Resources...

...wanted to measure expression of your protein of interest in a single cell? Or perhaps, you need to analyze...fluidics system they pass through a flow cell which restricts the size of the stream and forces the cells to...? However, fusion proteins tag your protein of interest with a fluorescent reporter on the N- or C-terminus... around the specific population that they are interested in and further analyze that subset of cells. .... An immunologist, for example, might be interested in the lymphocytes and “gate” that population for ...

...dabbles in science policy, public outreach, and restorative justice.  Follow her on twitter @sung_vanessa...age live in a state with at least two abortion restrictions that conflict with scientific evidence. We need...Campus.  He studies viruses including HIV and is interested in global health policy.  He is also chair of...Michigan, where programs like the Great Lakes Restoration Initiative are at risk of major cuts and where...science and positively reinforce their scientific interests. Our marches across the world have had an impact...

...parts in a specific orientation using Type IIS restriction enzymes (Weber et al., 2011). It can be performed...units (TUs) used to express your favorite gene-of-interest, all the way to the complex multi-TU constructs...and tags makes EcoFlex a great option for those interested in recombinant protein purification and metabolic...looking to express high levels of their gene of interest in yeast should also take a look at the Yeast ...stacking, increasing production of a protein of interest, and improving the nutrient profiles within given...

...always been interested in the ways that I could change the world around me. My strong interest in the intersections... marketing internship at Addgene. Although my interests are very clear, I had no idea how to translate...would be a perfect way of exploring my different interests but also a way of challenging what I am used to...

...multi-color imaging experiments. Interested in epitope tags? Others may be interested in attaching epitope tags... fluorescent proteins (FPs) to a yeast gene of interest. These fluorescently tagged genes are expressed...tags to their genes of interest, allowing for easy capture and detection of proteins and complexes, without...

...heavy restrictions on what I could and couldn’t do as an intern. I expected that these restrictions would...choose what gene to focus on. The gene choice had restrictions. It couldn’t be a Covid gene, a toxin, or an...running on a time crunch and knew that if we were to restart the procedure we would not have enough time. During...

... students to network, find groups with common interests, and settle into university life. The fairs are...1996, Cambridge Biosoc’s goal has been to promote interest in the biological sciences amongst the student...meetups. The society hopes to help young scientists interested in this field excel and develop their career ...from, and mentor while pursuing your research interests. Scientific societies like those mentioned above...

...the first gRNA using restriction enzyme BbsI and the second gRNA using restriction enzyme BsaI. If you’... of gRNAs Golden Gate Assembly uses Type IIS restriction enzymes, which cleave outside of their recognition...These expression vectors each contain Type IIS restriction sites flanking the promoter-gRNA construct, but...oligonucleotides. These plasmids contain Type IIS restriction sites that flank the promoter-gRNA constructs...

...plasmid with your gene of interest, you would replace mCherry with your gene of interest. pBLADEONLY: This plasmid...a pBAD33-based plasmid expressing your gene of interest, all you have to do is co-transform pBLADEONLY...pBLADEONLY. That way, a previously cloned gene of interest, under the pBAD promoter, can be controlled with light...

...well-equipped. Many institutions are sincerely interested in connecting with the public in a meaningful...scientific background and bring with them established interests and areas of expertise. Perhaps the strongest ... disparate spheres of research. In my case, an interest in neuroscience was carried over and allowed to... new intellectual environment. Over time, this interest came to incorporate other facets which occupied...

...available. Aguilar is a basic cell biologist. He got interested in Lowe Syndrome initially because of studies...disease associated with the gene, it piqued their interest. They obtained cells from patients with the condition...of the Wistar Institute, says that the lab got interested in XP out of a desire to understand the early...diseases at the same time. The limitation of budget restrained us from costly functional analyses.  “We believe...

...recapitulate the natural biology of a cell type of interest better than immortalized lines derived from the...it’s much more difficult to introduce a gene of interest into primary cells, so most primary cell lines...cell penetrating peptide (CPP) to a protein of interest thereby giving it the capacity to enter cells....29(1):33-41. PubMed. Protein transduction: unrestricted delivery into all cells? Schwarze SR, Hruska...

...Relationships need nurturing. Perhaps you’ve met someone interesting at an event. Or, perhaps you have a colleague...or other information that may be helpful or of interest to them. Don’t send a long email. If you have ...with people by discussing non-work topics and interests. Offer help when you can, instead of just asking... with people you don’t like or who do not show interest in you. Network with the people that you want ...

...protein (BFP, GFP, or RFP) to the genomic loci of interest. Pairs of identical hairpins within one gRNA will...HaloTag with red fluorescent ligands to the loci of interest. Dual color detection In order to label two loci...sgRNA-Sirius-8XPP7) to recruit GFP to the loci of interest. To determine how the sgRNA-Sirius-8XPP7 and sgRNA-Sirius...that result in yellow fluorescence at the loci of interest, the team could visualize three loci and measure...

...standardized DNA constructs, but its use of Type IIS restriction enzymes results in scarless assembly. Finally...allowing fragments to be taken directly from restriction enzyme digests to DNA assembly. What makes it...containing regions of homology) cut from plasmids via restriction enzyme digestion, and used directly in DNA assembly...as easy as obtaining a plasmid with the gene of interest (search Addgene plasmids by gene here), designing...