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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...
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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 ... -
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... -
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... -
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 ... -
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... -
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... -
Technical Design of a Western Blot
TypeBlog Post... incubation temperature for your specific protein(s) of interest. Determining linear detection range ...assay or BCA assay. Be sure to aliquot your sample(s) before adding in your denaturing and/or loading buffers...that covers a size range appropriate for the target(s) and transfer method. Membranes The two membrane ...your standard curve, your standard, and your sample(s), as you can only compare samples to a standard curve...using a protein stain like Coomassie Blue or Ponceau S. It is far less prone to error than normalizing to... -
CRISPR Guide Update
TypeBlog Post...blog PRIME Editing CRISPR 101: Cas9 vs the Other Cas(s) Progress Towards a PAM-free CRISPR... -
Neurodegeneration Plasmid Collection
TypeCollection...TDP43 RRM-GFP sigma-S TARDBP CMV ALS Rajat Rohatgi 107799 pHBS931 TDP43 RRM-GFP F-S TARDBP CMV ALS Rajat...TDP43 RRM-GFP FYW-S TARDBP CMV ALS Rajat Rohatgi 118792 pHBS1385 TDP43 RRM-GFP VLIM-S TARDBP CMV ALS Rajat...pHBS1410 GFP-TDP43 F-S TARDBP CMV ALS Rajat Rohatgi 118800 pHBS1411 GFP-TDP43 FYW-S TARDBP CMV ALS Rajat...BFP-P2A-TDP43 F-S] TARDBP CMV ALS Rajat Rohatgi 118809 pHBS1404 [IBB-GFP-mCherry 3E]-[BFP-P2A-TDP43 VLIM-S] TARDBP...SUMO-TDP43 F-S-TEV-mCherry TARDBP T5 ALS Rajat Rohatgi 133322 pHBS1552 H14-SUMO-TDP43 FYW-S-TEV-mCherry...-TDP43 F-S] TARDBP CMV ALS Rajat Rohatgi 133329 pHBS1505 [IBB-GFP-mCherry3E]-[BFP-TDP43 FYW-S] TARDBP ...Rajat Rohatgi 107800 pHBS935 TDP43 RRM-GFP M-S TARDBP CMV ALS Rajat Rohatgi 107801 pHBS934 TDP43 RRM-GFP... -
Antibodies 101: Monoclonal Antibodies
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... -
New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor
TypeBlog Post...Proteins Find AAV Tools References Marvin, Jonathan S., et al. “An optimized fluorescent probe for visualizing...PubMed Central PMCID: PMC4469972 Marvin, Jonathan S., et al. “Stability, affinity and chromatic variants... -
Lentiviral Prep Service
TypeCollection...Cas9 Cut Blasticidin Expresses human codon-optimized S. pyogenes Cas9 protein and blasticidin resistance ... -
Viral Vectors 101: Viral Vector Elements
TypeBlog Post...making! References and Resources Kulanayake, S., Tikoo, S. K. Adenovirus Core Proteins: Structure and ... -
Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2
TypeBlog Post...Ásgrímsdóttir ES, Rahimi K, Gill KP, Frandsen S, Hvolbøl Buchholdt S, Chen M, Kjems J, Febbraro F, Denham M ... -
7 Tips to Secure a STEAM Internship This Summer
TypeBlog Post...question(s). This precautionary step increases the likelihood of acceptance to the internship(s) you are... -
Summer SciComm: Preprints
TypeBlog Post...https://doi.org/10.1101/2021.09.07.459259 Sarabipour S, Debat HJ, Emmott E, Burgess SJ, Schwessinger B, Hensel...10.1371/journal.pbio.3000151 Sever R, Roeder T, Hindle S, Sussman L, Black K-J, Argentine J, Manos W, Inglis... -
Antibodies 101: Selecting the Right Antibody
TypeBlog Post...controls as you get your assays up and running. Ed.'s note: More questions on antibody validation? Check...availability References Uhlen M, Bandrowski A, Carr S, Edwards A, Ellenberg J, Lundberg E, Rimm DL, Rodriguez... -
CasPEDIA: A Functional Classification of Cas Enzymes
TypeBlog Post... https://doi.org/10.32942/X2C31F Makarova, K. S., Wolf, Y. I., Iranzo, J., et al. (2019). Evolutionary...CRISPR experiment? CRISPR 101: Cas9 The Other Cas(s) The PAM Requirement and Expanding CRISPR Beyond ... -
Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins
TypeBlog Post...inhibit type II CRISPR systems including the popular S. pyogenes Cas9 (Rauch et al., 2017, Hynes et al., ...DNA. What’s great about CASANOVA? It works with the S. pyogenes Cas9, dCas9-effector fusions, and xCas9,...