...of the undesired portion, designed to amplify the rest of the vector sequence. Encode homology in the forward... longer oligos, which could be avoided using restriction enzyme cloning, were more prohibitive at the ... doesn’t matter if the DNA is PCR amplified, restriction digested or even a synthesized gene. PCR is the...IVA can be employed by cutting the vector with restriction enzymes and adding the corresponding homologous...the MRC Laboratory of Molecular Biology. He is interested in neuronal signal transduction, and uses electron...

...sample names and guide sequences, and ICE will do the rest. There are no parameters that need optimizing and...This score is a useful measure for those who are interested in understanding how many of the contributing...

...part time work, bills, (at times) a social life and rest. I need it just as much in my current role as Customer...Support and Operations Manager at Addgene and is interested in nonprofit management and international development...

...activation kinetics and high responsivity near the resting membrane potential, ASAP5 is a great new option...CLAP) with a flexible linker where a protein of interest can be inserted. B) Protein expression and labeling...studies of kinesins but for many other proteins of interest too! Find CLIP, CLAP, and SNAP plasmids here! ...

...AMA Managing time and energy between work and the rest of life is sometimes more of a struggle and sometimes...negotiating a demanding career in science with other interests and priorities, but many find it helpful to obtain...

... that (1) the sequence is unique compared to the rest of the genome and (2) the sequence is located just...

...where you are going and what you expect. For the rest of the chalk talk, as with your job talk, avoid ...and tissues is fundamental to development. I'm interested in how cells find their way to the correct positions...expert in [your research area]," – you should show interest in other faculty members’ research. Still a Grad...Campus of the Howard Hughes Medical Institute. His interests include the quality control of secretory proteins...

...conditions:  1) The sequence is unique compared to the rest of the genome.  2) The target is present immediately...Cas9-mediated frameshift mutations in the gene of interest. The genomic target for Cas9 and the introduction...site early in the coding region of the gene of interest. This gives you loads of sequences to select from...to find a SpCas9 compatible PAM at your site of interest, that is the most efficient option. If there is...repressors to the promoter region of the gene of interest. dCas9-based repressors are useful when knocking...

.... Overgrown cultures stimulate quiescence, the resting phase of the cell cycle; quiescent cells do not...establish cell lines that stably express a gene of interest. While the general approach for using lentivirus...the positively charged polycation, polybrene. Interestingly, one study found that alternative polycations...

...between cancer cells, therapeutic drugs, and the rest of the body than studies in other model organisms...level and spatiotemporal expression of a gene of interest. This type of control over gene expression is ...continuously improved upon since then. If you are interested in these systems check out Gerald Rubin’s lab...specific plasmids (L4440) that contain your gene of interest (Fire et al., 1998). The discovery of RNAi in ...sensor, GCaMP6 (Nguyen et al., 2015). If you are interested in C. elegans plasmids and resources check out...

... easily gate on the unlabeled cells of interest while the rest can be excluded, or “dumped”. From left...fluorophore-conjugated antibodies directly bind to epitopes of interest that are found in the extracellular space. The...like bulk lympho-/splenocytes, you might only be interested in one of the various cell populations; for example...selection: labeling markers that your cells of interest do not have, and selecting for the cells that ...quick method of negatively selecting your cells of interest is magnetic bead-activated cell sorting, or MACS...An effective strategy to sort out your cells of interest via FACS using negative selection is called dump...Intracellular labeling Oftentimes, a marker of interest is not located on the cell membrane but inside...

... the experiment, preventing contamination of the rest. Avoid long exposure of the cell culture to air ...

...out my window and I saw the Dorr Mountain with the rest of the skyline of Acadia National Park. I was confused...molecular biologist with Biogen and is particularly interested in applying CRISPR for mouse genome editing. ...Zoology, Chinese Academy of Sciences, and he is interested in using CRISPR to develop therapeutic applications...

...exhausting. Now is the time to develop strategies to rest and re-energize yourself, and to address all of ...better idea of how to leverage it to ask new and interesting questions in your own lab. 2. Seek multiple mentors...investigators. Discussions can reveal mutual scientific interests, complementary perspectives, a shared communication...Campus of the Howard Hughes Medical Institute. His interests include the quality control of secretory proteins...

...so we’ll focus on DNA transposons for much of the rest of this article. Class I Transposable elements (...transposase gene is provided in trans so that a gene of interest can be inserted between the transposon’s LTRs,...mini-transposon that contains the DNA cargo of interest flanked by ~50-150 bp transposon end sequences...

...regions are made from the Ig heavy chains, and the rest of the Fab regions are completed with the Ig light... of interest from among the mixture and to measure the relative amount of your protein of interest on ...scientists to target specific proteins they’re interested in. Once a protein is targeted with an antibody...primary antibody, which recognizes your protein of interest, and a secondary antibody, which recognizes the... A primary antibody recognizing the protein of interest is added, and will bind to that protein on the... antibodies are used to capture the protein of interest and detect its presence. Then, the amount of light...sample, you’ll want to extract your protein of interest from the mixture. Antibodies can do this, too!...

...plasmid. This is seen only in the uncut DNA; the rest of the digest appears normal and produces the expected...When performing restriction digests on plasmids at Addgene, we sometimes observe something odd in our...themselves cannot be used to detect multimers, since restriction sites in multimers usually repeat in tandem along...DNA without changing the size. As a result, a restriction digest of multimers will produce the same fragment...sample, you could try linearizing the plasmid via restriction digestion and then run the linearized plasmid...eBookAddgene's Molecular Biology ReferenceAddgene's Restriction Digest Protocol More resources on the Addgene...

...presenting cells (APCs) that would normally alert the rest of the immune system to the presence of cancer are... complicated and dynamic phenomenon that is interestingly responsible for tumor reduction as well as progression...TGFβ. T-regs are suppressive immune cells that restrict the production and activity of other effector ... engineering and cancer immunotherapy, and is interested in exploring new avenues in healthcare.   References...

... monoclonal cell lines by limiting dilution. The rest of the cells were frozen down. Figure 1: ...

...a sequence recognized by the restriction enzyme MstII. When a restriction digest with MstII and southern...than real-time PCR (4). Restriction mapping and modern sequencing Restriction mapping informs on DNA sequence...different restriction enzyme digests. Incubating a DNA sample of unknown sequence with restriction enzymes...sequence of interest to create a physical map of the DNA. Starting in the 1970s, restriction mapping was...has changed, restriction enzymes are certainly not dead yet! Labs still keep restriction enzymes on hand...three historical techniques: southern blotting, restriction mapping, and sequencing gels, as well as their... (2). Southern blotting, in conjunction with restriction digest analysis, is used to identify the presence...