...Impressions of NgAgo In particular, we need to thank Pooran Dewari who wrote this month’s most popular blog...NgAgo genome editing up and running in their labs. Pooran surveyed a CRISPR google forum to get a better ...CRISPR-based technique to generate C->T or G->A transitions in a target sequence in live cells. This technique...endonuclease (dCas9) fused to a deaminase to make the transitions at target sequences specified by gRNAs. Caveats...Epigenomics 5 Tips for Troubleshooting Viral Transductions Single Base Editing with CRISPR Cas9 Activators...

...ve also carved pumpkins (yes, Blugene made an appearance there too) and shared fascinatingly creepy science...algae cell is just the cell wall and cytoplasmic membrane. Vampirovibrio chlorellavorus will not grown in... the snail, the larvae reach the eyestalk and transform them into a pulsating display imitating a moving...grasshoppers and crickets. When it’s ready to transform into an aquatic adult, it forces its insect host...uninfected grasshoppers, including those linked to neurotransmitter activity and geotactic behavior (Biron et ...

...details how to transfect CRISPR plasmids into MEL cells by electroporation, sort your transfected cells via...CRISPR/Cas9, we began to explore the paired double-strand break (DSB) approach at a variety of loci. We were...following paired DSBs is inversions [3]. These rearranged segments may also be informative as to genomic...Res. 2010 Jan;20(1):81-89. PubMed. 2. Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara Y, Lin C, et al. ...

... origin of replication (which controls the host range and copy number of the plasmid) and typically include...for further study. Promoter Region Drives transcription of the target gene. Vital component for expression...cell types. Primer Binding Site A short single-stranded DNA sequence used as an initiation point for PCR...vector containing the newly inserted gene is transformed into bacterial cells and selectively grown on...biologists to broadly utilize plasmids across a wide range of applications. Some common plasmid types include...

...chemistry. The society currently has 352 members, ranging from undergrad all the way up to professors. Oxford...Oxford currently has around 700 members and hosts a range of scientific meetings for students to attend. It...society provides talks and career support for a range of different scientific disciplines, as well as ... opportunities for society members as well as arranging collaborations and conferences for their students...Synthetic Biology Society also aims to develop a range of talks for both students and the general public...

...how much easier it makes our lives. Plasmid transformation into E. coli is a fairly inefficient process...aminoglycoside Binds 30S ribosomal subunit; causes mis-translation Bactericidal 50-100 ug/mL Spectinomycin aminoglycoside...Blocks 50S ribosomal subunit; inhibits aminoacyl translocation Bacteriostatic 50-100 ug/mL in EtOH Polymyxin...Polymyxin B polypeptide Alters outer membrane permeability Bactericidal 10-100 ug/mL Tetracycline tetracyclin...Binds 50S ribosomal subunit; inhibits peptidyl translocation Bacteriostatic 5-25 ug/mL in EtOH *In prokaryotes...

...sub-pathways like synthesis-dependent strand annealing (SDSA) and single-strand DNA incorporation (ssDI). "Easi-CRISPR...These complementary goals are aimed at improving translation of CRISPR technology, but they would also bring...a DNA repair newbie - diagrams of resection and strand invasion are like a foreign language to me. But...supports over 250 genomes, and they just released a brand new CRISPOR manual to kick off 2018! "Mechanisms...

...encompasses millions of choices including pharma, tech transfer, management consulting, science communication,...into industry.” In his article On Leaving Academe, Terran Lane is asked by his colleagues and friends why...tell you to pick up the pace Constant worry about grants and renewals What is different about non-academia...outsource a task someone else who can do better, no grant writing Job changes can happen regularly (every ...where you want to go will help you focus on the transferable skills that you will need to get there. Additional...

...reprogrammed into iEPs by retroviral expression of transcription factors Foxa1 and Hnf4α, a process that like...reprogramming (CellTagMEF), at the end of the three-day transcription factor reprogramming treatment (CellTagD3), ...determined by at least day 13. However, distinct transcriptional states were seen as early as day 6. All together...MettI7a1 encodes an uncharacterized putative methyltransferase. Adding Mettl7a1 to the standard Foxa1-Hnf4α...tagging two cell populations in a competitive transplant assay in the mouse intestine, demonstrating its...

...recombination, one resulting plasmid carries the transgene of interest (aka the viral genome) and the other...resistance gene and origin will replicate, and the transgene will be lost. Over time, bacteria containing the... grow! Our 5 Tips for Troubleshooting Viral Transductions will help with the next steps.   References...recombination of plasmid DNA without affecting mRNA translation efficiency or half-life. RNA 25:507–518. https...common lab E. coli strains Find tips for viral transduction  ...

...scientific community. Addgene was awarded a Fast Grant to ensure that the scientific community will continue...-2. The resurgence in popularity of the hACE2 transgenic mouse strain, for example, is sustained by the...University of Iowa (McCray et al., 2006), the hACE2 transgenic mice were designed after it was discovered that...Deposits of new reagents, policies from journals and granting agencies requiring materials to be deposited into... implications for clinical manifestations, transmissibility, and laboratory studies of COVID-19: an observational...

...with viruses or transmissible diseases, so we were left with no choice but to quarantine in our houses,...simpler and the volunteers helped with things like translations, bibliographic searches, etc.     Expanding...we call “asymmetric collaborations.” These are transient partnerships where a researcher makes a small ...blog post about remote collaborations Read our Transferable Skills Guide article about external collaborations...

...from green to orange such that non-growing cells are orange, slow growing cells are orange-green, and fast... and in assays for measuring human serotonin transporter activity. Unger also plans to use the serotonin...choice is the Arabidopsis thaliana - Pantoea agglomerans interaction. But the shape and metabolism of ...

...loci. 3. Cleave target DNA resulting in a double-strand break (DSB) Cas9 and its variants have two endonuclease...positions the nuclease domains to cleave opposite strands of the target DNA. Thus, the end result of Cas9...arrays as found in the bacterial genomes are transcribed into pre-crRNAs containing both the spacer region...RNase III, the tracrRNA and Cas9, bind to these transcripts and C) cleave them leaving mature crRNAs bound...target DNA which is E) cleaved leaving a F) double-strand break. A “gRNA” is a researcher-designed hybrid...

... different cell types so it is useful in a wide range of applications. This plasmid was the top requested...pSpCas9(BB)). The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP (PX458). lentiCRISPR...lentiCRISPR v2 - This 3rd generation lentiviral transfer vector also from the Zhang lab is used to deliver... David Root lab is a 3rd generation lentiviral transfer plasmid for cloning and expressing new shRNA sequences...the necessary elements for packaging, reverse transcription, and integration needed for the production of...

...alone. A few years ago when I started my career transition from bench science to software product management...explaining more of her role, which eventually led me to transfer to her team and begin my product management (PM...science backgrounds, most did not. It was my job to translate complex lab operations workflows into a format... help improve their work. Then, I distill and translate that valuable information back to the rest of ...science companies, try to find internships with translatable skills: working with data, working with people...

...microscopy, flow cytometry, or spectrophotometry. The range of fluorophores available for antibody conjugation... choice for ELISAs.  Biotin Biotin is used in a range of antibody-based assays. Biotin itself is not visible...Endogenous carbohydrates added to antibodies post-translationally. 4) Modifications made to antibodies, such ...commercially conjugated antibodies available, that never guarantees the antibody you need will be conjugated, or ...Stephenson W, et al (2017) Simultaneous epitope and transcriptome measurement in single cells. Nat Methods 14:...

...Signal transduction pathways are a lot like cell phone networks. Protein kinases deliver messages to ..., and how long their signals last all impact transduction of the message. To study signaling pathways,... game of telephone, often the signal must be transmitted by intermediate messengers. Optical control of...signaling by using psMEK1 and an ERK kinase translocation reporter with a mRuby2 fluorescent reporter ...feedback loops are a common feature of signal transduction pathways, but are often hard to decipher because...

... Also, many people on LinkedIn don’t appreciate random requests without a personal message and they may...trolls. People don’t like to connect with complete strangers as it borders on my first rule of networking--...a useful way to build or use a network. Total strangers can’t really help or refer you. Do not ask people... lot about requesting connections to complete strangers for informational interviews. Many have had success...considered intrusive and it’s still connecting with a stranger so is highly discouraged. You should be using ...

...Ambika, et al. "Transforming training to reflect the workforce." Science Translational Medicine (2015)...better solution would be to reduce the number of entrants into PhD programs to more closely reflect the ...symposium. Eve Marder, professor of Biology at Brandeis University, has recently published an article ...Among other involvements, he helped to establish grants through the Sloan foundation for postdoc-related...