...our live examples: experiment or analysis. In the rest of this article, we'll explain the thinking behind...

...is the right way?  First of all, I want to put to rest a common misconception. I find personal bias often...If you're interested in sharing your science, one of the big questions you'll be faced with is "how?"...be able to access your content.  What are you interested in? Math, sarcasm, romance, and spacecraft...can guarantee you that either resource or time restrictions (or both!) will be a major limiting factor at... to reach people who are not already actively interested in scicomm. Difficult to do in-depth storytelling...

...minutes of the two hour time slot, so she can take the rest of the time to collect her data. Perhaps you both...

...genome scale, it's going to be tricky to tame the rest of a cell. The BIOFAB plasmids represent the first...

...you even wear a lab coat! These are the things the rest of the world imagines when they think “scientist...your job as well as possible. It’s in their best interest to do so, sure, but it also eliminates that feeling...

...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...

...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...

...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...

...protein of interest in E. coli , B. subtilis and P. pastoris . All plasmids use the same restriction enzymes...relatively well understood biology to answer many interesting questions. Does my transcription factor bind ...and/or alter cellular activity in a way that’s interesting from an applied perspective, sometimes it’s best...Plasmids Tn7 Transposition Nancy Craig A gene of interest cloned into one of these plasmids can be transposed... many enterobacterial genomes. The bacteria of interest are transformed with the appropriate construct...get a better understanding of what your gene of interest does when expressed in vivo . Plasmid ID Visualization...fluorescent protein fusions with your protein of interest. See an overview of Addgene fluorescent protein...

...protein of interest. Signal strength is determined by how many antibodies bind the protein of interest, while... antibody of interest to a second, independent antibody for the same protein of interest, with no overlapping...quantify, identify, separate, and locate proteins of interest. The protein that an antibody recognizes is called... of the animal’s immune system. The antigen of interest is introduced to the animal, which then mounts...primary antibody which is bound to your protein of interest. Secondary antibodies can be isotype-specific ...recognize an available epitope on your protein of interest. Polyclonal antibodies are typically collected...Western blots should be run with your protein of interest and at least one ubiquitously expressed protein...

... of interest fused to the N-terminus of Amber Amber C1 Yellow Mammalian Express a gene of interest fused...a fluorescent fusion protein with your gene of interest and the listed fluorescent protein. Plasmid Color... mCerulean N1 Cyan Mammalian Express a gene of interest fused to the N-terminus of monomeric Cerulean ... mCerulean C1 Cyan Mammalian Express a gene of interest fused to the C-terminus of monomeric Cerulean ...His tag SCFP3A Cyan Mammalian Express a gene of interest fused to the C-terminus of SCFP3A Amber N1 Yellow...Amber mVenus N1 Yellow Mammalian Express a gene of interest fused to the N-terminus of monomeric Venus mVenus...mVenus C1 Yellow Mammalian Express a gene of interest fused to the C-terminus of monomeric Venus pCEP4YPet-MAMM...

... or by RT-PCR. Isolate your PCR product from the rest of the PCR reaction using a kit, such as the QIAquick...assist with restriction enzyme digestion (usually 3-6bp) Restriction Site: Your chosen restriction site for...fusing the upstream restriction site to the forward primer and the downstream restriction site to the reverse...product is now ready for restriction digestion. Digest your DNA: Set up restriction digests for your PCR ... Plasmid Cloning by PCR You may also like... Restriction Digest of Plasmid DNA DNA Ligation Bacterial ...of a piece of DNA and at the same time adding restriction sites to the ends of that piece of DNA so that...that it can be easily cloned into a plasmid of interest. For this example, we will describe how to copy...

...-CDS issues. Prohibited Restriction Sites: EcoRI, XbaI, SpeI, PstI Restriction Sites to Avoid: NotI For...Coli genome. Prohibited Restriction Sites: EcoRI, SpeI, NheI, PstI Restriction Sites to Avoid: PvuII, ... use of assembly standards – combinations of restriction sites which preserve a standard prefix and suffix...backbone can be repeated to ligate your gene of interest after the promoter, and so on. The following table... reverse complimented for primer design. The restriction enzymes used for assembly are the prohibited ... Employs high-efficiency enzymes. Prohibited Restriction Sites: EcoRI, BglII, BamHI, XhoI For more info...giving in-frame scar encoding Thr-Ala. Prohibited Restriction Sites: EcoRI, XbaI, SpeI, PstI For more info,...

...Experimental Tips Choosing a tet system If your gene of interest should be active, and only turned off occasionally...off or tTA is more appropriate. If your gene of interest should be mostly inactive, and only turned on ...tetracycline-dependent promoter upstream of your gene of interest tTA expression plasmid Components for a tetracycline...tetracycline-dependent promoter upstream of your gene of interest rtTA or TetR expression plasmid Stable cell lines...will allow you to insert and induce your gene of interest. ID Plasmid Description Co-expressed tTA, rtTA... 16542 pBI-MCS-EGFP Expression of your gene of interest (MCS with a β-globin poly A) & EGFP from a bidirectional...plasmid for inducible expression of transgene of interest and EGFP None Either Lung 11662 pPRIME-TET-GFP-FF3...

...analysis to purify specific genomic regions of interest 48807 8xLexA-binding elements/pMD20 iChIP plasmid...analysis to purify specific genomic regions of interest 51258 3xFLAG-dCas9/pMXs-IG Expresses 3xFLAG-dCas9...analysis to purify specific genomic regions of interest 51259 3xFLAG-dCas9/pMXs-I2 Expresses 3xFLAG-dCas9...analysis to purify specific genomic regions of interest 51260 3xFLAG-dCas9/pMXs-neo Expresses 3xFLAG-dCas9...analysis to purify specific genomic regions of interest 61079 gRNA-hIRF-1 #12 Expresses a guide RNA (gRNA...analysis to purify specific genomic regions of interest 63589 3xFN-Tel-TAL/pCMV-7.1 Expresses a 3xFLAG-tagged...analysis to purify specific genomic regions of interest 82613 3xFLAG-dCas9/MSCV-EGFP Expresses 3xFLAG-...

...can precisely control expression of your gene of interest. Many Cre constructs also contain fluorescent ...a promoterless vector with several available restriction sites none Mammalian Sauer 11957 pBS595 tet-hCMV-EGFPcre...with stop codon Entry vector Ko 139546 pENTR-CreStopDeleted Cre without stop codon Entry vector Ko 139547...-lox” or “LSL” cassette) upstream of a gene of interest will prevent gene expression in the absence of... Switch: These constructs contain two genes of interest, Genes A and B. When Cre is absent, only Gene ...and type of target sites that flox your genes of interest makes FLEx switch a powerful experimental tool...information. Empty backbones to regulate your gene of interest: ID Plasmid Description Expression System PI 11578...

...Express gene of interest from the CMV promoter pCDH-EF1 72266 Express gene of interest from the EF-1 promoter... Express gene of interest from the CB promoter pCDH-PGK 72268 xpress gene of interest from the PGK (phosphoglycerate...Express gene of interest from the CB promoter pCDH-EF1s 72484 Express gene of interest from a truncated...respectively, and empty vectors to express your gene of interest under the control of a variety of promoters. Professor...kinase I) promoter pCDH-CMV4 72284 Express gene of interest from the CMV promoter. WPRE has been removed to...

... 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...