...mean you have one copy of the gene; an RQ value of 2 means you have two copies, and so on and so forth....quantitative PCR. Expert Rev Mol Diagn. 2005 Mar;5(2):209-19. doi: 10.1586/14737159.5.2.209. PMID: 15833050...to a reference sequence present in the same amount in all samples (usually to a ubiquitously expressed,... reference sample with two copies of the gene (Ma, 2014.) As with all qPCR assays, samples, standard curves...housekeeping’ gene), and quantified relative to a 'normal' sample with known copy number set as the expression baseline...since you’re looking at the amount of DNA in your sample relative to your normal control - you can set up...number (RQ value) matches the copy number of your sample. In that case, an RQ value of one (or, more realistically...

...is unique compared to the rest of the genome and (2) the sequence is located just upstream of a Protospacer...pyogenes Cas9 variants with varying PAM sequences, 2) the use of Cas9 homologs derived from species other...Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell 163:759–771 . https://doi....and crRNA (or synthetic gRNA) are derived from the same species. Expanding the CRISPR toolbox The isolation...

...China, Korea, Japan, Brazil, India, and Mexico. Table 2: Addgene distributors Distributor Partnership...distribution after we partnered with them (Figure 2). In others, the distributor helped solve an issue...temperature for the duration of their journey. Figure 2: Annual distribution of items to countries with a ...that available and accessible weren’t always the same thing, shaped both Addgene’s founding and our growth...

...FRET signal back again. The Ting lab constructed 2 versions of this reporter to target the K9 (pcDNA3... 525.7570 (2015): 533-537. PubMed PMID: 26352475. 2. Bygren, Lars O., et al. "Change in paternal grandmothers...corresponding gene products due to the same region, but not the same exact genes, being improperly silenced.... This is how most cells in the body contain the same DNA but have very different structure and function...present without any DNA changes, and both involve the same region of chromosome 15. They both result from the...

...target sites for editing. PAM sequences can be from 2-6 base pairs in length and follow the DNA sequence... Eric Perkins Mammalian genomic DNA is not just a 2-D linear molecule snaking through our cells’ nuclei...SpCas9) requires an NGG PAM to recognize the target. Osamu Nureki’s lab engineered SpCas9 (SpCas9-NG) with ...

...episode 2 "Organizing Literature" : I found out about Zotero here Dear Grad Student podcast episode 2 "Essential...and Linux.  Mendeley and Zotero have a lot of the same features, but this chart highlights the differences...

...the reporter mRNA is labeled by PCP-mCherry (Figure 2). The 3’ UTR also contains a CAAX sequence to bind...PMC2746483 Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An RNA biosensor...highly useful, this technique requires extensive sample processing to avoid disrupting ribosome-mRNA complexes...

...immediate early gene expression. Genes Dev. 1988 Jun;2(6):718-29. doi: 10.1101/gad.2.6.718. PMID: 2843425...Targeted Gene Expression in Drosophila.” Genetics 186(2) (2010): 735-55. PubMed PMID: 20697123. PubMed Central... in Saccharomyces cerevisiae.” FEMS Yeast Res. 12(2) (2012): 197-214. PubMed PMID: 22129153. Additional...system to GAL4/UAS that functions in essentially the same manner, with LexA binding to the lexA operator (...desirable to control or look at multiple genes at the same time.  The 3 binary systems listed above are orthogonal...

...label) your sample (i.e. cultured cells or tissues).  Step 2: attach anchors to your sample that will link...so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC protocols and you might...we just made the sample bigger? In ExM, and its various derivatives, biological samples are enmeshed in...the number of specific targets you can label in a sample. Super-resolution microscopy (SRM), a family of... its original size. Depending on the protocol, samples can be expanded to ~4-10x linearly! This expansion...gel components to polymerize. Step 3: digest the sample to allow for even expansion.  In some cases, labels... ExM process allows you to physically magnify a sample so you don’t have to optically magnify quite so...

...green squares, purple circles and blue triangles. 2) An antibody immobilized on a bead binds specifically...protein G, a novel IgG-binding reagent. J Immunol 133(2):969-974. Hjelm H, Hjelm K, Sjöquist J (1972) Protein... adjusted depending on the sample. It is important to remember that sample lysates will also contain proteases..., incubate the sample as you would in an IP using a nonspecific antibody from the same host species as... from the sample prior to beginning the IP.  During the IP, the target protein in the sample binds to ...parallel with both a sample known to express the protein of interest, such as a sample transiently transfected...out in one of two ways. Both methods utilize the same basic strategy: immobilize the antibody or antibody-target...

...linear fashion. This implies that reducing the power 2-fold does not reduce photobleaching by half. How exactly...prolonged time). Other illumination strategies, such as 2-photon-excitation, selective plane illumination, TIRF...This can differ between fluorescent proteins in the same spectral class (Cranfill et al, 2016). This non-...comparison of several fluorescent proteins of the same spectral class on the microscopy system and in the...

...11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016...isolated which target and cleave RNA through the same mechanism as traditional spCas9. If you are looking...Cas13 and has been proposed to be useful for the same applications (RNA knockdown, RNA editing, etc.)....

...that the signal to noise ratio can be increased from 2-fold (by CRISPRainbow) to 3-fold with CRISPR-Sirius...CNV) analysis of the U2OS cells.   Figure 2: CRISPR-Sirius tricolor labeling. (A) Diagram of sgRNA-Sirius...combination of MS2, PP7, or boxB sequences in the same sgRNA will generate additional colors. Image from...

... you can read more about them in review articles (2–4). You can also take a look at my recent paper to...Development 118, 401–15 (1993). PubMed PMID: 8223268. 2. J. B. Duffy, GAL4 System in Drosophila : A Fly Geneticist...this mutant fly to one lacking the mutation in the same genetic background. In other cases, YFG or one of...

... Zetsche et al., Nat Biotechnol. 2015 Feb 2;33(2):139-42. doi: 10.1038/nbt.3149. Light-inducible ...and on your favourite signalling pathways (Figure 2). They are all available now at Addgene. 1 Lungu...20;19(4):507-17. 10.1016/j.chembiol.2012.02.006. 2 Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; ...Nihongaki et al., Chemistry and Biology. 2015 Feb 19; 22(2):169-74. High efficiency cloning for enzyme screening...inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene deletion, (3) simultaneous...different fluorescent proteins, 8 constitutive promoters, 2 includible promoters, 3 polyA terminators and various...of plasmids required for each input/output system; 2) removing all chemically inducible promoters; 3) improving...

...scientists have 1) the courage to make mistakes and 2) the integrity to recognize these mistakes quickly...She studies beta-cell function and failure in Type 2 Diabetes. Birmingham, AL, USA 33.5207°N 86.8025°W ...available to people who want to join the dialog and 2) remember that science should not be partisan: everyone...campus speakers thoughtfully reminded us to use the same critical lens on our scientific community as we ... nor would you mention my small error and at the same time pass over in silence his very large one” (http...exoplanets into a successful PhD thesis. We use the same atmospheric physics to understand exoplanets that...happy to see so many others come out who feel the same.  I was also incredibly impressed with how well ...

...expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...cell 58.4 (2015): 644-659. PubMed PMID: 26000849. 2. Grimm, Jonathan B., et al. "A general method to improve...removing excess dye from the sample, which is particularly important for samples where washing is difficult...reason is something called “the photon budget”: each sample has a limited number of fluorophores and each fluorophore...of information one can extract from a biological sample is wholly dependent on the photon budget and pushing... desperate to extract a few more photons from a sample. Chemical fluorophores can be substantially brighter...

...Table 2: Lab strains of E. coli Strain Natural resistance Primary Use Genotype ccdB Survival 2 T1R ...resistance to tetracycline Additionally, Table 2 provides a quick reference for some of the popular...convoluted due to researchers sharing and renaming samples throughout history. It was likely isolated in 1918...

...sites would confound your experimental results and 2) perform targeted off-target measurements at those...experiments where only localization of the Cas occurs. 2 gRNA approach Off-targets that come from mutations...perform downstream characterization of, isolating 2 or 3 distinct clones to perform key experiments on...If you are performing CRISPR library screens, the same principle holds true - the higher the number of ...

..., and the desired gene (transfer vector plasmid), 2) the HIV-1 genome containing all viral genes with ...protocols that have been described (protocol 1, protocol 2, protocol 3) to find the one that will give you the...PubMed PMID: 3785217. PubMed Central PMCID: PMC367857. 2. Naldini, Luigi, et al. "In vivo gene delivery and...are working with proto-oncogenes or with human samples that have not been tested for HIV.  If not using...ribosome entry site (IRES) and expressed under the same promoter. Alternatively, your gene and the marker...