...into a vector with a large tag at the N-terminus, but you don’t have the tag already included in the vector...you can insert both the gene of interest and the tag sequences into the vector in one step without scars...In this blog post, I will go over some advantages, disadvantages, and examples of how scientists are using... common, and for good reason. They offer many advantages over the traditional restriction enzyme cloning...thereof) nor a serious time commitment. Another advantage is that this process makes it easy to generate...primer melting temperature greater than 48℃. Both Snapgene and NEB have tools that help you design primers...video gives a helpful demonstration of how to use Snapgene’s program to design primers for Gibson Assembly...

...plasmid.     Confirming tags and fusion proteins Finally, we confirm promoters, tags, fusion proteins, and...aligning to NCBI entry or using BLAST.   Confirming tags and fusion proteins. We’ll break down each of these...selectable markers by detecting common features using Snapgene. If we find information that differs from what...

...short sequences of up to 100 bp, such as epitope tags, design primers with template binding regions to...combining multiple primers, a C-terminal epitope tag can be moved to the N-terminus of a protein-coding... can be introduced in a gene, while adding a GFP-tag (Figure 3). Achieving multiple modifications requires... to the 5’ end of the reverse primer. Mutagenesis - Mutagenesis is performed by designing primers astride...modification, from insertions, deletions, and point mutagenesis to complex assembly of multiple genes. Primer...a sequence manager A sequence manager, such as SnapGene, can be very helpful to confirm primer design....rapid method for recombination and site-specific mutagenesis by placing homologous ends on DNA using polymerase...

...to an epitope tag or fusion protein. If your sequencing primer was chosen to confirm a tag or fusion protein...expected ORFs, mutations or truncations. A primary advantage of blastx is that you do not have to decide on...recommend using blastp over blastx. The primary advantage is time savings but an added benefit is that blastp...other matrices to see if another would be more advantageous for your search. Timesaving Tip #3: Note that...at NGS Plasmid Quality Control Learn about our Snapgene-powered plasmid maps. Resources on Addgene.org...

... Many thanks to our guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at the Janelia...Growth of new livers in culture could resolve this shortage." What is the knowledge gap you plan to address...

... Many thanks to our guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at theJanelia... be needed to achieve your intended next career stage and create a timeline for achieving these goals....

...Academic Job Interview by Erik Snapp Addgene depositor and guest blogger Erik Snapp breaks down how to prepare... 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins by Cary...

...contains 24 copies of SunTag, a synthetic scaffold that can recruit GFP fused to the SunTag-specific antibody...biochemical method developed in 2009, takes a “snapshot” of all of the mRNA bound-ribosomes in a cell....Imaging by Coat protein Knock-off) instead takes advantage of the ribosome’s movement. When the ribosome ...indicating translational repression. During later stages, single RFP-labeled osk and Oskar protein were ...scFv. Each time the reporter is translated, the SunTag sites on the nascent polypeptide recruit scFV-GFP..., as the authors of these papers did. Since the SunTag-based system tethers mRNAs to the plasma membrane...Additional Resources the Addgene Blog Learn More about SunTag Catch up on Your GFP Background Choose the Brightest...

...newly synthesized product from a site-directed mutagenesis experiment? The answer to both questions is the...to methylation if the enzyme's recognition site (TCTAGA) is preceded by GA or followed by TC. As shown ...cleave DNA. DpnI is often used for site directed mutagenesis. During this process, incorporation of a desired...illustrates how sequence visualization software (we used Snapgene here) can help you assess whether your restriction...Troubleshoot Your Sequencing Results Use REPLACR Mutagenesis to Mutate Your Plasmids Resources on Addgene.org...

...three larval stages where larvae molt in order to continue growing in the next larval stage. After the ...the third larval stage, Drosophila will pupate, which takes about one week from the initial deposition of...through their translucent bodies. This is the ideal stage to select for females because you can be certain...Drosophila cell lines are extremely effective. Their Snapdragon tool helps you select primers to synthesize RNA...

...and in more than 30 countries around the world. Snapshots of the BioBuilder experience With its intentionally...recognized platform in March to exhibit projects at any stage of completion, from pitch to prototype. Last year...preparing local (Massachusetts), economically disadvantaged students for paid summer internships. A summer...

...more about the Gateway cloning method and its advantages. An introduction to Gateway technology The Gateway..., etc). This video demonstrates how to use the Snapgene program to design Gateway plasmids.    ...and verify that your construct is functional. Advantages of the Gateway cloning method Compatibility and...JY, Chin CF (2015) Gateway Cloning Technology: Advantages and Drawbacks. Clon Transgen 4:138. doi:10.4172...

...possible consequences of FP aggregation, as Erik Snapp’s blog post shows. To avoid these issues, Subach ...knowledge of protein structure and saturation mutagenesis. mK-GO matures from green to red, and the FT ...absorption spectra). Subach et al’s directed mutagenesis also produced the three separate timers shown...

...lineage, without dedifferentiating into a pluripotent stage. Cellular reprogramming The reversal of a differentiated...reprogramming [1]. The process can be divided into two stages: Dedifferentiation - Conversion of adult somatic...differentiated cells of choice. The dedifferentiation stage involves overexpression of four reprogramming factors...differentiated somatic cell to revert back to a pluripotent stage (iPSC formation) [2, 3]. The iPSCs then proliferate...reprogrammed to iPSCs successfully [10-12]. The next stage of reprogramming consists of redifferentiation of... resources More (due to additional pluripotency stage) Less Safety Less (C-MYC, a known oncogene is ...Williams, L.A., B.N. Davis-Dusenbery, and K.C. Eggan, SnapShot: directed differentiation of pluripotent stem ...

...grows on a daily basis, the lists below provide a snapshot of relevant resources and a good jumping off point...H3.1 H3.3 HIST1H1E HIST1H2BB HIST2H2BE RBX1 STAG2 Xist Your favorite gene! SWI/SNF Genes:       ...

...tendency – mean, median, and mode give you a summary snapshot without telling you anything about outliers or...dependent variables worked synergistically, antagonistically, were additive, etc.). ANCOVA and MANCOVA...