...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...

...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...

...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...

...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!...

...best-characterized effects of p53 are in promoting cell cycle arrest, apoptosis, or senescence in damaged cells. The... receptor Gadd45 GADD45A GADD45B GADD45G Growth arrest and DNA-damage-inducible; alpha, beta, or gamma...BCL2 binding component 3 Reprimo TP53 dependent G2 arrest mediator candidate Scotin Shisa family member 5...

...promoter or regulatory regions of your gene of interest. If the plasmid that you choose does not also ...transcription. Design your gRNA to target your gene of interest’s promoter/enhancer or the beginning of the coding...specific promoter or enhancer for your gene of interest. Available modifications include histone acetylation...

...encode a wide range of gene products. The main restrictions on a lentiviral transfer plasmid are size (i.e... Any expression cassette (promoter and gene of interest) can be cloned into the plasmid. Nolan 12257 pWPXL...bi-cistronic expression of EGFP and the gene of interest, also can express shRNA (cloning H1-shRNA cassettes...

...plasmids for endogenous tagging of your gene of interest.... design. How to use CRISPR to tag your gene of interest Mendenhall and Myers Tagging System The Eric Mendenhall...plasmids as listed in each row. If your own gene of interest is currently unavailable, you will need to design...

...conditions: The sequence is unique compared to the rest of the genome The target is present immediately ...Publications CRISPR Overview Bacteria have an interesting adaptive immune response that detects and eliminates... Cas enzyme and a gRNA specific to the gene of interest. The genomic target of the gRNA can be any ~20...activity with endogenous cell-type specific miRNAs to restrict CRISPR editing only to the target tissue. Browse... molecules associated with a genomic region of interest in vivo. Figure 12: Overview of purifying genomic...

...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: ...

... molecules associated with a genomic region of interest in vivo is essential to understanding locus function... molecules associated with a genomic region of interest in vivo. For CRISPR-based purification, design...

...localization and/or function of your protein of interest by fusing it to a fluorescent protein. FRET Find...Subcellular Localization Determine where your protein of interest resides by using a well-characterized fluorescent...

...repeat arrays of any desired length: the REAL (Restriction Enzyme And Ligation) assembly method (Sander ...Targeter enables users to search their sequences of interest for potential TALEN or TALE transcriptional activator...

...the system is to infect the cell population of interest with a deletion-mutant tracing virus that lacks..., Young JA, Callaway EM. 2007b. Monosynaptic restriction of transsynaptic tracing from single, genetically...

...promoter or regulatory regions of your gene of interest. If the plasmid that you choose does not also ...transcription. Design your gRNA to target your gene of interest’s promoter/enhancer or the beginning of the coding...

...microbiology resources: Find plasmids for your microbe of interest, including bacteria, viruses, protozoa, fungi,... cerevisiae - Dueber Lab Yeast Prototrophy : Restoration of prototrophy in common S. cerevisiae lab strains...

...promoter or regulatory regions of your gene of interest. If the plasmid that you choose does not also ...transcription. Design your gRNA to target your gene of interest’s promoter/enhancer or the beginning of the coding...

...for plasmids containing your zebrafish gene of interest? Search Addgene's collection for plasmids that...vertebrate brain - Alex Schier Lab Other Tools of Interest A few examples of popular tools to use in zebrafish...

...promoter or regulatory regions of your gene of interest. If the plasmid that you choose does not also ...transcription. Design your gRNA to target your gene of interest’s promoter/enhancer or the beginning of the coding...