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TypeBlog Post... because I only left him/her with small tasks and s/he was always asking for more work (this is a big ...your network…. I found it helpful to plan my week(s) in advance as much as possible so I could more easily...support you, just like you are supporting your report(s). This means that you should be able to go to your...
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To Codon Optimize or Not: That is the Question
TypeBlog Post...codon bias across all endogenous genes in E. coli and S. cerevisiae (Tuller et. al., 2010). Figure ...doi.org/10.1038/nsmb1031 Gustafsson C, Govindarajan S, Minshull J (2004) Codon bias and heterologous protein...synthetic biology in Natural or synthetic? How Addgene;s dataset reveals trends in biological innovation Resources... -
A Look Back at One Year of Plasmid Sharing for COVID-19 Research
TypeBlog Post...deposited plasmids for the expression of cathepsin S, L, and B. This became a great starting point for ...Bertozzi CR, Carette JE, Stanley SA, Harris E, Konermann S, Hsu PD (2021) Genome-wide, bidirectional CRISPR screens...Garcia N, Whiteman MB, McKay LGA, Lelis FJN, Habibi S, Cai Y, Rennick LJ, Duprex WP, McCarthy KR, Lavine... -
Hot Plasmids: Winter 2025
TypeBlog Post... Hao, Y. A., Lee, S., Roth, R. H., Natale, S., Gomez, L., Taxidis, J., O'Neill, P. S., Villette, V., Bradley... -
Tips for Screening with Yeast Two Hybrid Systems
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... -
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... -
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... -
Trimmer Lab NeuroMab Collection
TypeCollection...190500 Homer1L/S scFv [L113/130] L113/130 scFv 2a Homer1L/S Mouse Mouse 190501 Homer1L/S scFv [L113/130...Gamma2L/S scFv [N452/73] N452/73 scFv 2T GABA(A)R, Gamma2L/S Human Mouse 190556 GABA(A)R, Gamma2L/S scFv ... Human Mouse IgG2a 177456 Anti-Homer1L/S [L113/130R] Homer1L/S Mouse Mouse IgG2a 177457 Anti-Homer1L [...IgG2a 177553 Anti-GABA(A)R, Gamma2L/S [N452/73R] GABA(A)R, Gamma2L/S Human Mouse IgG2a 177554 Anti-GABA...N476/9R] ZIP3 Mouse Mouse IgG2a 206600 Anti-S-tag [N56/9R] S-tag Bovine Mouse IgG2a 206601 Anti-TrpC4 [...Human Mouse IgG2b 206636 Anti-Homer1L/S [L113/130R-2b] Homer1L/S Mouse Mouse IgG2b 206637 Anti-Homer1L... Human Mouse IgG1 206687 Anti-Homer1L/S [L113/130R-1] Homer1L/S Mouse Mouse IgG2a 206688 Anti-Homer1L ... -
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... -
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... -
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 ... -
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...