...solution or something in between, you are using a tag with a serious drawback. The standard EGFP plasmid... of a POI. As an investigator, you want a fusion tag to be inert, to not produce artifacts in your experiments...Many thanks to our guest blogger Erik L. Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by guest blogger Erik L. Snapp. Stop using EGFP/GFP for fusion proteins! Despite...oligomers such as DsRed or high affinity FPs such as TagRFP. Problem 3 can be solved, in most cases, by using...distance runner, gardner, and cook.   References 1. Snapp, E.L., et al., Formation of stacked ER cisternae... Resources on the Addgene Blog Read Professor Snapp's Post on mOX FPs Find the Best Fluorescent Protein...

...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...

...categories of biosensors or browse all the biosensors tagged in our catalog . Use the article links to find ...2023.07.18.549527 Eric Schreiter Calcium GCEPIA1-SNAP ER for sensing ER calcium A genetically encoded ...by a novel reporter protein, tandem fluorescent-tagged LC3. Autophagy. 2007;3(5):452-60. Tamotsu Yoshimori...Super-Ecliptic, pHluorin-mKate2, Tandem Fluorescent Protein-Tagged Human LC3 for the Monitoring of Mammalian Autophagy...Hoffman Voltage JEDI-2P voltage indicator for two-photon imaging Sustained deep-tissue voltage recording...Rafael Yuste Voltage Ultrafast fluorescent voltage sensor ASAP2f Subcellular Imaging of Voltage and Calcium...fluorescent voltage sensor. Nat Neurosci. 2014 Apr 22. Michael Lin Voltage Fluorescent voltage reporters...

...Includes tagging with CFP, YFP, and mRFP Tsoulfas Lab Lentiviral Plasmids - Fluorescently tag your gene...empty plasmid backbones with different fluorescent tags for you to create fusion proteins with your gene... iRFP Gradia Lab Mammalian Plasmids - Includes tagging with mCherry, mCitrine, mCerulean Davidson Lab ...C. elegans Hamdoun Lab Plasmids - Set includes tagging with mCherry, Cerulean, Citrine, and eGFP pPD95... expression Yeast Thorn Lab Vectors - Includes tagging with a wide variety of colors, some specifically...Lindquist Lab Vectors - Gateway cloning; includes tagging with ECFP, EYFP, DsRed, Cerulean Bacteria Gradia...

...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...

...Plasmids encoding fluorescent proteins tagged with genes or peptides with known subcellular localization...Voeltz 79802 pTag-RFP-C-h-Rab5a-c-Myc Early endosomes Rab5a TagRFP James Johnson 79801 pTag-BFP-C-h-Rab5a-c-Myc...Mothes 79806 pTag-RFP-C-h-Rab11a-c-Myc Recycling endosomes Rab11a TagRFP James Johnson 79805 pTag-BFP-C-h-...endosomes Rab11a TagBFP James Johnson 79800 pTag-RFP-C-h-Rab4a-c-Myc Recycling endosomes Rab4a TagRFP James Johnson...EEA1 GFP Silvia Corvera 42635 TagRFP-T-EEA1 Early endosomes EEA1 TagRFP-T Silvia Corvera 38770 pEF.myc.ER-E2...moxGFP Erik Snapp 68126 ERoxBFP Endoplasmic Reticulum ER retention signal oxBFP Erik Snapp 49150 BFP-KDEL...Johnson 79799 pTag-BFP-C-h-Rab4a-c-Myc Recycling endosomes Rab4a TagBFP James Johnson 12674 GFP-rab11 ...

... Pages. Addgene sequences the plasmid to verify tags, mutations and a portion of the insert, but we do...that can open .ab1 files, such as 4Peaks (Mac), SnapGene Viewer (Mac/PC), FinchTV (Mac/PC), Sequence Scanner...

.../9R] ZIP3 Mouse Mouse IgG2a 206600 Anti-S-tag [N56/9R] S-tag Bovine Mouse IgG2a 206601 Anti-TrpC4 [N77...channel Mouse Mouse IgG2a 140071 Anti-Snapin/SNAPAP [L8/15R] Snapin/SNAPAP Human Mouse IgG2a 140072 Anti-Mitofusin... Human Mouse IgG2a 114476 Anti-Synaptotagmin-3 [N278/19R] Synaptotagmin-3 Mouse Mouse IgG2a 114477 Anti-TRPML3... Human Mouse IgG2a 114482 Anti-Synaptotagmin-12 [N277/7R] Synaptotagmin-12 Mouse Mouse IgG2a 114483 Anti-GluA1...) Rat Mouse IgG2a 114558 Anti-Synaptotagmin-10 [N269/73R] Synaptotagmin-10 Mouse Mouse IgG2a 114559 Anti-Ataxin... Mouse Mouse IgG2a 177526 Anti-Synaptotagmin-7 [N275/14R] Synaptotagmin-7 Mouse Mouse IgG2a 177527 Anti-Lgi1... Human Mouse IgG2a 188216 Anti-Synaptotagmin-6 [N270/47R] Synaptotagmin-6 Mouse Mouse IgG2a 188217 Anti-CASK...

... 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 ...