...postdocs in science, engineering and health from the NSF’s 2014 Graduate Student Survey data). Most institutions...populations potentially from 1 to 5761, according to the NSF data) and therefore the numbers of postdocs at different...implies. These 5 organizations, according to the NSF data, account for 18% of the postdoctoral workforce...

...reduce barriers to implementation. Funded by the NSF (with Co-PIs Jessica Bell, Lisa Gentile, Ellis Bell...Hampshire. He mentors undergraduates conducting NIH- and NSF-funded research on planarian flatworms both in and...). But is it possible to bring the kinds of transformative educational experiences afforded to those fortunate...NADH production.   CUREs are beginning to transform undergraduate education in the sciences by making...

...Improving your work-life balance Balancing the Scale: NSF's Career Life-Balance Initiative Can Scientists Maintain...

...STEM education is supplemented. A 2009 study by the NSF showed that among US 4th graders tested on the Science...

...Carefully transfer the transfection mix to the Lenti-X 293T packaging cells. Add the transfection mix dropwise...general method for transfecting mammalian cells using linear polyethylenimine. Transfections allow for transient...lentiviral vectors) Day 1 (pm): Transfect Cells Day 2 (am): 18h post transfection - Remove media, replace with...protocol to transfect mammalian cells with your plasmid of interest. Protocols... Protocols General Transfection General...General Transfection You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral Service...adapted for different cell lines and different transfection reagents. Last Upload: June 10, 2016 Workflow...

...Introduction Transformation is the process by which foreign DNA is introduced into a cell. Transformation of bacteria...prepared for optimal transformation efficiencies upon thawing. For the highest transformation efficiency, we...time when carrying out transformations? If you are not concerned with transformation efficiency (such as ... 20-30 mins) I get very few if any transformants when transforming large plasmids (>10 kb) or BACs, what...Learn how to transform E. coli with your plasmid of interest. Protocols... Protocols Bacterial Transformation Bacterial...Bacterial Transformation You may also like... Plasmid Cloning by Restriction Enzyme Digest Restriction Digest...

... to proceed with transfection. Transfect the flask containing 108 mL cells: Transfer 6 mL of BCD TFX into...General Transfection Protocol Recombinant Antibody Purification Protocol Introduction Transfections allow...using Polyethylenimine Max as a transfection reagent. After transfection and expression, the recombinant...Section 2: Transfection Check the cell density and viability: Using a 5 mL pipette, transfer 0.5 mL of ...feeding schedule: Thursday: Transfect cells. Friday (24 h post-transfection): Add 4% BCD Feed. Monday (...post-transfection): Add valproic acid to 3.75 mM, add 4% BCD Feed. Tuesday (120 h post-transfection): Add...Wednesday (144 h post-transfection): Add 4% BCD Feed. Thursday (168 h post-transfection): Harvest antibody...

...pOpen-EcoRIM M.EcoRI DNA methyltransferase 165517 pOpen-Eco31IA Eco31IA DNA methyltransferase 165518 pOpen-NotIM...pOpen-NotIM M.NotI DNA methyltransferase 165527 pOpen-Eco31IB Eco31IB DNA methyltransferase 165536 pOpen-PstIM...pOpen-PstIM M.PstI DNA methyltransferase 165538 pOpen-SfiIM M.SfiI 165538 DNA methyltransferase 165547 pOpen-SapIM1...pOpen-SapIM1 M1.SapI DNA methyltransferase 165557 pOpen-SapIM2 M2.SapI DNA methyltransferase 165558 pOpen-EcoRVM...pOpen-EcoRVM M.EcoRV DNA methyltransferase 165577 pOpen-XbaIM M.XbaI DNA methyltransferase 165578 pOpen-NcoIM ...into an expression vector of their choice and transforming into E. coli bacteria. This can help reduce ...an expression vector of their choice and then transform into E. coli bacteria for expression and purification...

...another transfection method such as cationic liposome-based transfection. Optimize transfection conditions...ligation to occur in one reaction (see step 3.2 ). Transform 10 μl of DH5α E. coli cells with 1 μl of reaction...Perform maxi-prep for each CRISPR/Cas9 construct. Transfecting CRISPRs into Cells of Interest NOTE: This protocol...which is used for MEL cells. However, transient transfection of CRISPR/Cas9 plasmids may be successfully ...types using preferred culture conditions and transfection strategies for each cell type. While MEL cells... before attempting genome editing. Immediately transfer solution from cuvette into 1 ml of culture media...Minimize the time between electroporation and transferring the solution into media to enhance cell viability...

...Addgene lentiviral packaging, envelope, and transfer plasmids...Libraries All Viral Plasmids Packaging Plasmids Transfer Plasmids Lentiviruses, a subset of retroviruses...encodes the viral genome (typically called the transfer plasmid ). This table lists common lentiviral ... Contains Rev; can be used with 3rd generation transfer plasmids along with additional packaging components...Contains Gag and Pol; can be used with 3rd generation transfer plasmids along with a plasmid encoding Rev Trono...and Tat; can be used with 2nd and 3rd generation transfer plasmids Trono 12263 pCMV delta R8.2 2nd Packaging...and Tat; can be used with 2nd and 3rd generation transfer plasmids Trono 12259 pMD2.G N/A Envelope VSV-G-expressing...

...lentiviral and adenoviral transfer vectors with Addgene. Lentiviral Transfer Vectors Within this set of...packaging these transfer vectors and generating infectious virus. Protocols: Lentivirus Transfer Vector Production...Core Lentiviral and Adenoviral Vectors Lentiviral Transfer Vectors Adenoviral Vectors You may also like..... of lentiviral transfer vectors you can find reporter plasmids to test whether or not you’ve efficiently...coexpressed with tdTomato and Pac (puromycin N-acethyl-transferase) pCDH-CB-copGFP-T2A-iCre 72256 Expresses copGFP...PGK promoter Adenoviral Vectors The adenoviral transfer vectors contained within this set are derived ...Flp recombinases. Genes can be cloned into these transfer vectors using shuttle vectors pICPIS-EF1 and pICPIS-CB...

...positively transfected. Alternatively, you can use a selection marker to select transfected cells (for..., the most efficient and easiest way is to co-transfect several plasmid together, with each plasmid having...two spacers for each locus into PX330, then co-transfect or delivery the two plasmids together into your...run them and then select the best guide to co-transfect in HR donor. Alternatively, a new system described...oligo is fine based on new data). For general transfection, you could use 1ul of 10uM oligo stock for each... or mutation. Each arms is around 800bp. For transfection in cells like HEK, we use around 400ng of a ...single-point mutation, the best HR template for transfection is probably a single stranded DNA (ssDNA) oligo...

... latent transforming growth factor beta binding protein 1 MGC163161 LTBP2 latent transforming growth factor...thyroglobulin AITD3, TGN TGFA transforming growth factor, alpha TFGA TGFB1 transforming growth factor, beta 1 ...TGFbeta TGFB2 transforming growth factor, beta 2 MGC116892, TGF-beta2 TGFB3 transforming growth factor...containing) transforming protein 2 SCK, SHCB, SLI SHC3 SHC (Src homology 2 domain containing) transforming protein...MGC138230 TGFB1 transforming growth factor, beta 1 CED, DPD1, TGFB, TGFbeta TGFB2 transforming growth factor...TGF-beta2 TGFB3 transforming growth factor, beta 3 ARVD, FLJ16571, TGF-beta3 TGFBR1 transforming growth factor...protein 2 C14orf141, LTBP3, MSTP031 LTBP3 latent transforming growth factor beta binding protein 3 DKFZp586M2123...

...For easy-to-transfect cell types (e.g. HEK293 cells), transfection with standard transfection reagents may...be present in a single lentiviral transfer vector or separate transfer vectors May contain reporter gene...-target activity. By screening E. coli cells transformed with a pooled library of SpCas9 variants, researchers...libraries) are a heterogenous population of lentiviral transfer vectors, each containing an individual gRNA targeting...then cloned in a pooled format into lentiviral transfer vectors (panel B ). CRISPR libraries have been...pattern opens up the possibility of directional gene transfer, analogous to traditional restriction enzyme cloning...or gRNA in a mammalian cell line that can be transfected at high efficiency Lentiviral transduction Cas9...

...constitutive promoter and can be used to monitor transfection or viral infection efficiency, to study changes...complementation in plants. Bioluminescence Resonance Energy Transfer (BRET) CalFlux VNT : An intracellular calcium ...white screening can be used for screening of transformants in cloning process. Lynne Postovit 119760 pSGDlucV3.0... Renilla luciferase is used to monitor viral transfection Feng Zhang 21471 pLenti PGK V5-LUC Neo (w623... Firefly luciferase is used to moniter viral transfection Sidi Chen 109396 TBG-Vector Firefly TBG AAV-... Firefly luciferase is used to moniter viral transfection Sidi Chen 118412 ssAAV-EF1a-FLuc-WPRE-HgHpA_...

... Material Transfer Agreements Most Addgene orders must be accompanied by a material transfer agreement...agreement (MTA), including transfers to for-profit entities. The MTA is a legal agreement between the recipient... the depositing institution(s). It governs the transfer of biological materials and intellectual property...Please see the Sample Agreements on our Technology Transfer Information page for more information. To learn...

...expression vectors, human embryonic kidney cells are transfected with the plasmid(s) encoding the heavy and light... The fluorescence pattern is compared to an untransfected control. Immunohistochemistry (IHC) Tissue sections...extracted from cells, separated by SDS-PAGE, transferred to a membrane, and labeled with the antibody ...antibody. Protein expression is compared to an untransfected control and the protein size is confirmed. Residual...contain low levels of co-purified plasmid from transfection. This residual plasmid is extracted and amplified...

...Kasparov Popular γ-Retroviral Transfer Plasmids Common γ-retroviral transfer plasmids. Want a retroviral...Addgene, including popular packaging, envelope, and transfer plasmids. Viral...of interest? Search our database for retroviral transfer plasmids that have inserts. ID Plasmid 5'LTR Description...

...design of synthetic operons for chloroplast transformation and metabolic engineering in plants (including...set of functional parts for assembling plant transformation vectors - Daniel Voytas. Transcription Factors...Agrobacterium-mediated transient expression or plant transformation - Silin Zhong. RNAi The following collections...Editors Save Time with Transient Plant Leaf Transformations RNA Interference in Plant Biology: New Tools...

...resonance energy transfer, or FRET , is a process by which energy is non-radiatively transferred from an excited...excited donor fluorophore to an acceptor. Since the transfer of energy does not occur by emission of a photon...FRET experiments. The efficiency of the energy transfer is measured using fluorescence microscopy by exciting...