...Samples of viral preparations are subjected to polyacrylamide gel electrophoresis (PAGE) followed by silver...centrifugation and then by filtration through a 0.45 µm polyethersulfone (PES) membrane. The preparations are then ... 293T cell line in the presence of 10.0 μg/mL polybrene. This ensures that any loss of titer associated...

...civilization in general (1). Science and politics? Science is an apolitical process for seeking knowledge. The...inclusion of science to inform policy (17). The facts surrounding a matter or policy should have more weight ...science rallies scientists to political action, and inspires politicians and citizens to include science... again. No part of this process is inherently political so why is there a march in Washington, D.C., the... the seat of the United States government? Politics and science are intrinsically linked: governments ...when we drop them. Scientific realities limit what policy can do and, in this way, science has always been...been related to government and, consequently, politics. More concretely, scientists gather information ...

...vaccinia virus - Bennink and Yewdell Labs Yarrowia lipolytica Golden Gate tool kit : Integrative vector assembly...transcription units in the oleaginous yeast Y. lipolytica - Rossignol Lab Yeast Gateway kit : 285 vectors...

...Adeno X shuttle vector with EF1 promoter and SV40 polyA pICPIS-CB 73356 AdenoX shuttle vector with chicken-beta...chicken-beta actin CMV enhancer promoter SV40 polyA...

... own RNA-dependent RNA polymerase into the infected cells. The EBOV polymerase replicates the viral genome...under control of a T7 RNA polymerase promoter sequence. Thus, T7 RNA polymerase also needs to be present...gene start) and a transcription termination/polyadenylation signal sequence (gene end). A minigenome is...replication and reporter gene transcription by the EBOV polymerase (reviewed in 2, 5). The minigenome is capable...nucleoprotein, NP. NP associates with the viral polymerase, which is formed by a complex of two proteins...sense, single-stranded RNA minigenome. The viral polymerase complex then recognizes the encapsidated minigenome... order to replicate the minigenome, the viral polymerase initiates RNA synthesis at the 3’ end of the ...

...conditions. Expression of an error prone polymerase I (PolI3M) is capable of mutagenesis in E. Coli, but... error prone polymerase to introduce mutations. However, by using nCas9 to direct PolI3m to a particular...Figure 1: EvolvR consists of an error-prone polymerase (PolI) that is fused to a nicking Cas9. A guide ...EvolvR: (1) two additional mutations to PolI3M generated PolI5M which increased EvolvR’s mutagenesis rate... from bacteriophage T7 DNA polymerase increased the processivity of PolI3M, and thereby increasing the... system has two key components: the error prone PolI3M that’s fused to a nicking Cas9 (nCas9). Similar... site of interest. A gRNA is used to direct the PolI3M-nCas9 complex to a DNA site of interest, which ...

...approach policy makers take in abusing climate science skepticism to promote harmful policies that benefit...move slowly, news about possible broad changes to policies seems to break everyday. Many researchers and ...participation in government, evidence to inform policies, a protected place to work, funding for science...medicines, technologies, defense, and governing policies are developed primarily by wishful thinking. The...rains, it was weather befitting of the ominous political climate in which American scientists find themselves...scientists found themselves not too long ago.   From policies that “muzzle” government scientists to the de-...Canada.  Outside the lab, she dabbles in science policy, public outreach, and restorative justice.  Follow...

...Sharing Policy Discussion PLOS’ New Data Policy: Public Access to Data PLOS’ New Data Policy: Part Two...advice about the scope of the policy. End of story? Not quite. The PLOS policy on data access is a formal...clarified what the policy intent was and provided an FAQ list to further explain the policy. The fact that...year when it announced its data access and sharing policy. Since early March, the open access publisher has...the research reported in PLOS publications. The policy was not an overnight revelation, rather it was ...reasonable. PLOS has since clarified their data sharing policy and recently announced that of the 16,000 manuscripts...agencies (e.g. Wellcome Trust, NIH Data Sharing Policies, etc.) and is founded on the principle that scientific...

...purification based on the polyhistidine tag. What is the polyhistidine tag? The polyhistidine tag is a string of...      Fig. 1: A sample containing a polyhistidine tagged protein of interest (purple) and several...to the Ni2+ and is immobilized along with a polyhistidine containing contaminating protein (orange). A...is successfully isolated.   Prior to the polyhistidine tag, alternative fusion tags were used, such...interfering with protein function. In contrast, the polyhistidine tag is very small and rarely affects protein...function (Bornhorst 2000).  One downside of polyhistidine tag-based purification, however, is the propensity... containing Co2+, have a lower affinity for polyhistidine than more commonly used substrates such as Ni2...

...AAV-mediated transduction and targeting of retinal bipolar cells with improved mGluR6 promoters in rodents...Intravitreal delivery of a novel AAV vector targets ON bipolar cells and restores visual function in a mouse model...

...2 like 2 NFKB1 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 PAK PAK1 PAK2 PAK3...containing protein 1 SCRIB Scribbled planar cell polarity protein SHC SHC1 SHC2 SHC3 SHC4 SHC (Src homology...

...you need a high-fidelity polymerase. There are many high fidelty polymerases on the market; you need one...a DNA polymerase that produces blunt-ended PCR products (e.g. Phusion, Pfu, and Vent polymerases). Note... PCR followed by Restriction Fragment Length Polymorphism (RFLP) analysis (right). Introduction of restriction...amplified fairly easily by simply following the polymerase manufacturers’ PCR protocol. Be sure to adjust...plasmid template for site directed mutagenesis.  Polymerase To ensure that you don’t introduce undesired ... one with 5’->3’ polymerase activity (for amplification), 3’->5’ exonuclease activity (increases amplification...Note that certain polymerases such as Taq generate A-overhangs (a feature used in TA-cloning). Such non-complementary...

...have huntingtin proteins with abnormally long polyglutamine sequences. These glutamine-rich sequences are...environmental factors. For example, variations of Apolipoprotein E (APOE), such as the ε4 allele, are a risk...

... Science. 2017 Nov 3;358(6363):623-630. Franck Polleux Calcium Red calcium sensor PinkyCaMP based on mScarlet...Encoded Glutamate Indicators with Altered Color and Topology. ACS Chem Biol. 2018 Jan 22. Robert Campbell Glutamate...exploiting a temperature-responsive elastin-like polypeptide. Sci Rep. 2021 Aug 13;11(1):16519. Takeharu Nagai...

...window) reverse transcriptase, protease, and Bst polymerase constructs now available from Andrea Pauli's ...Diagnostic Toolkit Enzymes. (unpublished) Taq polymerase and MMLV reverse transcriptase expression plasmids...

...      The RNA polymerase(s) RNA is transcribed from DNA using an RNA polymerase (RNAP). In bacteria...bacteriophage Constitutive, but requires SP6 RNA polymerase. SP6 polymerase has a high processivity. When used for...describes the combination of the promoter (RNA polymerase binding site) and operators (response elements...promoter region controls the binding of the RNA polymerase and transcription factors, therefore promoters...single enzyme; however, eukaryotes have muliple polymerases which are each responsible for a specific subset... used in expression systems for Drosophila. Polyhedrin General expression mRNA  Strong insect promoter... H1 small RNA expression shRNA From the human polymerase III RNA promoter Constitutive  May have slightly...

...path to science policy and science communication A career in science communication or policy requires broad... space of science education, communication, and policy. In this post, I hope to tell you a little more...technologies and as societal participants in the political process. My role at pgEd I have always been very...attending seminars in bioethics, public health, science policy, and science and technology studies (STS). I also...genetics research, their applications and regulatory policies; keeping tabs on the conversation about genetics...posts, lesson plans, or information briefs for policymakers. The rapidly changing landscape of genetics ...seminars, symposia, or courses in social, health or policy-related topics, as time permits. Also seek out ...

...CaMKII-somBiPOLES-mCerulean CamKII Bidirectional Optogenetics Wiegert 154951 hSyn-DIO-somBiPOLES-mCerulean...

... bodies are a polymer of many copies of monomeric subunits. Unlike cytoskeletal polymers, however, R bodies...This post was contributed by guest blogger Jessica Polka, a Postdoctoral Research Fellow with Pamela Silver...force by dynamically rearranging into different polymerization states. Instead, R body monomers respond to...engineering objectives are possible in our recent paper (Polka and Silver 2016). First, we demonstrated that R ...mechanically with their environment.   Jessica Polka is a Postdoctoral Research Fellow with Pamela Silver...Harvard Medical School. Follow her on Twitter @jessicapolka.     References 1. Kanabrocki, J. A., Quackenbush...Protistology 48, 290–296 (2012). PubMed PMID: 22356923. 3. Polka, J. K. & Silver, P. A. A Tunable Protein Piston ...

...with a 6x His tag. Requires a strain with T7 RNA polymerase. Amp resistant. More expression vectors for protein... of the pB1H2w derivatives as a fusion to RNA polymerase subunit ω. Transcription factor binding to a ...