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TypeBlog Post... heterologous systems (e.g. mammalian proteins in S. cerevisiae). While this may be difficult to resolve...1985; 43:729-736. PubMed PMID: 3907859. 4. Fields, S., Song, O. A novel genetic system to detect protein-protein...membrane protein interactions. Proc Natl Acad Sci U S A. 2004; 101(33):12242–12247. Pubmed PMID: 15299147...
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RNA Interference in Plant Biology: New Tools for an Old Favorite
TypeBlog Post...processed miRNA or siRNAs then silence the target gene(s) either transcriptionally or post-transcriptionally...produced that contains sequence against your gene(s) of interest and the reporter. This approach greatly...Plant Biology 7:251–257 . https://doi.org/10.1055/s-2005-837597 Li J-F, Chung HS, Niu Y, Bush J, McCormack...F, Joberty G, Zinn N, Mueller WF, Clauder-Münster S, Eberhard D, Fälth Savitski M, Grandi P, Jakob P, ... -
A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression
TypeBlog Post... and resources References Aschauer DF, Kreuz S, Rumpel S (2013) Analysis of Transduction Efficiency, Tropism... OS, Velardo MJ, Peden CS, Williams P, Zolotukhin S, Reier PJ, Mandel RJ, Muzyczka N (2004) Recombinant....org/10.1016/j.ymthe.2004.05.024 Dudek AM, Pillay S, Puschnik AS, Nagamine CM, Cheng F, Qiu J, Carette... -
Antibodies 101: Introduction to Antibodies
TypeBlog Post...012198382-6/50026-1 Wang R, Xiang S, Feng Y, Srinivas S, Zhang Y, Lin M, Wang S (2013) Engineering production...doi.org/10.3389/fimmu.2017.01589 Ayyar BV, Arora S, Ravi SS (2017) Optimizing antibody expression: The... -
CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More
TypeBlog Post...detectable using PCR. The system also functions in S. pneumoniae and can be used to generate multiple mutations...gene lambda Red. pTargetF contains the specific gRNA(s), and the repair template is supplied as a dsDNA fragment...Perutka J, Powell JE, Geng P, Richhart DD, Byrom M, Kar S, Davies BW, Ellington AD, Moran NA, Barrick JE (2018....7b00399 Li Q, Sun B, Chen J, Zhang Y, Jiang Y, Yang S (2021) A modified pCas/pTargetF system for CRISPR-...AK, Carette X, Potluri L-P, Sharp JD, Xu R, Prisic S, Husson RN (2016) Investigating essential gene function... -
Isolating a Monoclonal Cell Population by Limiting Dilution
TypeProtocol... in the number of integration events and the site(s) of proviral integration across cells. Selective pressure... -
Affinity Purification of Recombinant Antibodies with Protein A or Protein G
TypeProtocol...hydrochloride pH 9.0. Cap the tubes and vortex for 5 s to mix. Determine the protein concentration of each... -
Sensing Neuronal Dopamine
TypeBlog Post...showed rapid response (60 or 130 ms on and 0.7 or 2.5 s off, for mouse or human receptors, respectively). ...AAV vector inventory. References Darvesh, Altaf S., et al. "In vivo brain microdialysis: advances in... -
Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation
TypeBlog Post...research community. References 1. Coroadinha, A. S., et al. "Effect of osmotic pressure on the production...bioprocesses (1989): 272-278. 4. Ozturk, Sadettin S., and Bernhard O. Palsson. "Effect of medium osmolarity... -
Antibodies 101: Affinity Tags
TypeBlog Post...western blot and immunofluorescence. GST Glutathione S-Transferase (GST) is a 26 kDa tag which can be isolated...doi.org/10.1002/0471140864 Fox, J. D., & Waugh, D. S. (2002). Maltose-Binding Protein as a Solubility Enhancer... -
Viral Vectors
TypeCollection...'s Biosafety Resource Guide . References Cohen S, Au S, Panté N. How viruses access the nucleus. Biochim... -
Deep Mutational Scanning with One Pot Saturation Mutagenesis
TypeBlog Post...this. 2) When your gene of interest has a BbvCI site(s). If so, the nicking mutagenesis protocol will still...still work if the orientation of the BbvCI site(s) of the gene are in the same orientation as those in ...site directed mutagenesis to remove the BbvCI site(s). Overall, the key features of the one-pot saturation... -
Antibodies 101: Normalization and Loading Controls for Western Blots
TypeBlog Post...between protein abundance and band intensity is an S-shaped curve. Sparse proteins may not be detected .... Created with BioRender.com. To see why this S-shaped relationship poses problems for proteins of... of this gel is misleading. We can see why on the S-shaped curve. On the protein abundance axis, both ... -
CRISPR 101: Validating Your Genome Edit
TypeBlog Post...Renaud J-B, Schneider-Maunoury S, Shkumatava A, Teboul L, Kent J, Joly J-S, Concordet J-P. Evaluation of...e58. https://doi.org/10.1093/nar/gky164 Cancellieri S, Canver MC, Bombieri N, Giugno R, Pinello L. CRISPRitz... -
Viral Vectors 101: Pseudotyping
TypeBlog Post...//doi.org/10.3390/v12091016 Joglekar AV, Sandoval S (2017) Pseudotyped Lentiviral Vectors: One Vector,...10.1128/jvi.71.8.6174-6178.1997 Wickersham IR, Finke S, Conzelmann K-K, Callaway EM (2006) Retrograde neuronal...Wickersham IR, Lyon DC, Barnard RJO, Mori T, Finke S, Conzelmann K-K, Young JAT, Callaway EM (2007) Monosynaptic... -
Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping
TypeBlog Post...vivo (Ge et al., 2019). This technique relies on an S. auerus transpeptidase sortase A variant (mgSrtA),...-1837. PubMed PMID: 30676735. Kim, D. I., Jensen, S. C., Noble, K. A., KC, B., Roux, K. H., Motamedchaboki... PubMed Central PMCID: PMC4831873. Lam, Stephanie S., et al. "Directed evolution of APEX2 for electron... -
Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View
TypeBlog Post...to invert or ‘flip DNA’, is derived from the yeast S. cerevisiae (Gronostajski and Sadowski 1985). The ...References 1. Atasoy, D., Y. Aponte, H. H. Su, and S. M. Sternson. 2008. “A FLEX Switch Targets Channelrhodopsin...Ligand-Dependent Chimeric Cre Recombinase.” Proc Natl Acad Sci U S A 92(15): 6991–95. PubMed PMID: 7624356. PubMed Central... -
Addgene Receives NIH BRAIN Initiative Grant to Create Open-Access Recombinant Antibody Resource
TypeBlog Post...Trinklein ND, Thie H, Wilkinson IC, Tandon AK, Anderson S, Bladen CL, Jones B, Aldred SF, Bestagno M, Burrone... XY, Eng H-Y, Lane D, Lefranc M-P, Clark M, Dübel S (2018) When monoclonal antibodies are not monospecific... -
Revamp Your Lab Meetings With Creative Virtual Collaboration
TypeBlog Post...Denfeld BA, de Eyto E, Hampton SE, Keller PS, Sharma S, Lewis ASL, Weyhenmeyer GA, O’Reilly CM, Lofton ME...https://doi.org/10.1371/journal.pcbi.0030044 Wuchty S, Jones BF, Uzzi B (2007) The Increasing Dominance ... -
Choosing Your Fluorescent Proteins for Multi-Color Imaging
TypeBlog Post...that can be imaged with the optics on the microscope(s) you intend to use. An accurate determination of whether...protein that is supposed to be extremely bright. In S. cerevisiae, we've tested a number of green and red...