...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 ...

...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...

...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...

...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...

...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...

...      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 ...

... comes from using Taq polymerase for the amplification step since Taq polymerase leaves a single deoxyadenosine...mind that Taq polymerase has an error rate of about 1 in 3,500 bases. Typically polymerases with proofreading...and amplify your sequence of interest with Taq polymerase using your favorite PCR protocol.  2. Set Up ...to reduce error rates; however, proofreading polymerases will also remove all unpaired 3’ ends in your... 2. Novel approach to molecular cloning and polynucleotide synthesis using vaccinia DNA topoisomerase....

... DNA Polymerase I 165533 pOpen-TthpolA Tth DNA polymerase 165534 pOpen-T7gene5 T7 DNA Polymerase (unmodified... Ligases DNA Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases Other Enzymes... of molecular biology, including DNA polymerases, RNA polymerases, DNA ligases, reverse transcriptases... DNA Polymerases Addgene ID Plasmid Gene/insert 165500 pOpen-9N7polA 9 degrees N-7 DNA polymerase 165501.... Heat-Stable DNA Polymerase 165502 pOpen-9N7polA (CT) 9 degrees N-7 DNA Polymerase chain terminating ...pOpen-K12polLF DNA Polymerase I, Large (Klenow) Fragment 165522 pOpen-K12polLF (exo-) DNA Polymerase I, Large (...

...The key is T4 DNA polymerase, which has 3′→5′ exonuclease activity. T4’s polymerase activity can be thwarted...thwarted if no dNTPs are available to polymerize, allowing the slower exonuclease activity to proceed.... of the chosen nucleotide, T4 will switch to polymerizing and then stall due to the absence of the other...dNTP present in the exonuclease reaction or polymerization will occur at that position, preventing T4 ...target vector. 5′ overhangs can be generated by T4 polymerase (left branch); by annealing two separate PCR ...advantage of SLIC over Gibson assembly is cost, as T4 polymerase is much less expensive than the enzymes required...

... protein mix by size Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) is the first...of a resolving gel composed of the crosslinked polymer acrylamide. In addition to the samples, a protein...membrane, usually composed of a nitrocellulose or polyvinylidene difluoride (PVDF) in a step called protein ...state while a western blot detects denatured polypeptide chains. The protein regions available for antibody...between a protein in its native state versus the polypeptide chain. Consequently, antibodies validated for...transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic...) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and...

...class I polypeptide-related sequence A FLJ60820, MGC111087, PERB11.1 MICB MHC class I polypeptide-related...catalytic, delta polypeptide p110D PIK3CG phosphoinositide-3-kinase, catalytic, gamma polypeptide PI3CG, PI3K...gonadotropin, beta polypeptide CGB3, hCGB CGB1 chorionic gonadotropin, beta polypeptide 1 - CGB2 chorionic... gonadotropin, beta polypeptide 2 - CGB5 chorionic gonadotropin, beta polypeptide 5 HCG, MGC119822 CGB7... beta polypeptide 7 CG-beta-a, FLJ35403, FLJ43118 CGB8 chorionic gonadotropin, beta polypeptide 8 - CHGA... - GIP gastric inhibitory polypeptide - GIPR gastric inhibitory polypeptide receptor MGC126722 GKN1 gastrokine...factor alpha polypeptide PDGF-A, PDGF1 PDGFB platelet-derived growth factor beta polypeptide (simian sarcoma...

...Single polycistronic lentiviral vector for the expression of human Oct4, Sox2, and Klf4 Polycistronic lentiviral...27(5):1042-9. Townes Lentivirus Human Single polycistronic lentiviral vector for the expression of human...(8):1221-9. Cibelli Lentivirus Human Single polycistronic, doxycycline-inducible lentiviral vector for...Non-integrating mini-intronic plasmid (MIP) polycistronic expression of human Oct4, Klf4, Sox2, c-Myc ...episome Human Non-integrating EBNA1-mediated polycistronic expression of human Sox2, KLF4, L-Myc, Lin28...Replicating EBNA1 episome Human Non-integrating polycistronic expression of human Oct4, Klf4, Sox2, c-Myc,...episome Human Non-integrating EBNA1-mediated polycistronic expression of human Oct4, Sox2, Myc, Klf4, BCL2L1...

...downstream manipulations. Phi29 polymerase is a phage-derived polymerase that can perform whole genome ... approaches a host cell surface are complex polysaccharide structures, many of which are easily varied...fragmentation, repair sheared ends with T4 DNA polymerase (to create blunt ends) and ligate into a bluntly...manipulation, amplifying phage DNA with phi29 polymerase prior to fragmentation can greatly facilitate...into E. coli BL21 (DE3) cells (these have the T7 polymerase and a few proteases knocked out making them particularly...

...to express the transcription factors from a polycistronic cassette. However, it was unknown whether variations...variations in gene expression from these polycistronic cassettes could lead to comparable experimental...Woltjen Lab compared different variants of the polycistronic cassettes and discovered reprogramming discrepancies...variation in the Klf4 isoform (KLF4s or KLF4L). Polycistronic cassettes that contained KLF4s had overall diminished...reprogramming and global gene expression patterns. Polycistronic cassettes that contained the KLF4L isoform had...the observed reprogramming discrepancies. The polycistronic piggyBac transposon vectors tested in this work.... They are compatible with both agarose and polyacrylamide gels. They are shared through Addgene without...

...lot number while you’re at it! Polyclonal or monoclonal Both polyclonal and monoclonal antibodies can ...can work well for a western blot. Polyclonal antibodies, which bind to multiple sites on an antigen, are...protein — can be used to reproduce the benefits of polyclonal antibodies. Because these mixtures are defined...are not as commonly available as monoclonal or polyclonal antibodies, but it’s worth a look to see if they...Precast gel Increased cost Save money Polyclonal antibodies Higher non-specific binding; lot-to-lot...available; cost varies widely Increase signal Polyclonal antibodies Higher non-specific binding; lot-to-lot...

...incubating Nanoblade particles with polybrene, a cationic polymer. Polybrene has long been used to increase...from Mangeot et al., 2019.   MLV GAG-POL polyprotein Nanoblades are formed when viral structural proteins...protease, supplied as part of the MLV Gag-Pol polyprotein, cleaves the proteolytic site connecting the ...

...negatively-charged membranes (Davis et al., 2002). Polybrene is one such cationic reagent that is commonly ...efficiency by 10-fold (Davis et al., 2002). However, polybrene is highly sensitive to freeze-thaw cycles and ...be stored in single-use aliquots. Never use a polybrene solution that has been thawed more than once. ...in cells that are sensitive to the toxicity of polybrene (e.g., hematopoietic or primary cells), and has...., Jeffrey R. Morgan, and Martin L. Yarmush. "Polybrene increases retrovirus gene transfer efficiency ...

...sufficient to seed polymer formation as fusion protein concentration increases. The polymerized fusion proteins...dimerizing FP has potential to form large stable polymers of dimers. 3) FRET biosensors that contain two...dimerization coupled with POI dimerization can lead to polymerization of the fusion protein. A dimeric fusion protein...resulting in false positives [2, 4]. Solutions to the polymer problem There are some simple solutions available...

...not suited to generate polycistronic containing multiple identical polyadenylation signals or promoters ...fluorescent proteins, promoters, enhancers, polyadenylation signals, inducible gene-expression sequences...promoter (to drive high-level expression) and a polyA signal (to terminate transcription) and the four...constructs were then combined to create a single polycistronic 15 kb insert. The final construct was then introduced...