...consensus sequence for the addition of a poly(A) tail, the addition of a poly(A) tail on a prokaryotic transcript...any 3’ regulatory elements, such as the polyadenylation or poly(A) signal. While many studies focus on ...eukaryotic polyadenylation signal directs cleavage at the cleave signal and addition of a poly-A tail to...in the RNA transcript followed by a weakly bound poly-uracil tract as shown in the figure to the right...eukaryotic-specific process, prokaryotes also add poly(A) tails to certain RNAs. Unlike the eukaryotic mechanism...added to any accessible 3' end. The presence of the poly(A) tail targets the RNA to the degradosome, which...protected by secondary structure. It is thought that poly(A)s are used to control the cellular concentration...

...makes polyclonal antibodies unique and what experiments you should choose them for. How polyclonal antibodies...cross-reactivity. When to use polyclonal antibodies One advantage of polyclonal antibodies is that they are...barrier to entry makes polyclonal antibodies a common reagent in many labs. Polyclonal antibodies also have...When to stay away from polyclonal antibodies The most prevalent issue with polyclonal antibodies is their...from the same species your polyclonal antibody was generated in or a polyclonal isotype control, to account...options to choose from. In this article we’ll cover polyclonal antibodies, one of the many different types of...antibodies are produced Polyclonal antibodies are a heterogeneous mixture of many antibodies that recognize...

...resistant DNA polymerase from the thermophilic bacterium, Thermos aquaticus (Taq). Taq polymerase is thus a...slightly to Taq polymerase’s ideal temperature (70-75oC). At this temperature Taq polymerase can synthesize...mutagenesis.  Quantitative polymerase chain reaction, qPCR Quantitative polymerase chain reaction (qPCR) also...What is polymerase chain reaction? If you have ever worked in a molecular biology laboratory you have...have likely done a polymerase chain reaction (PCR). PCR is an in vitro method in which a small amount of...Yellowstone National Park.  In addition to Taq DNA polymerase, PCR requires free nucleotides (dNTPs), template... Primers are crucial for this process as DNA polymerases require an existing strand of DNA to add nucleotides...

... science policy, there is often discussion of “big P” and “little p” policy.  “Big P” policy is what you...about where science policy happens.  When do you do science policy? Many science policy training programs... is science policy?”  I have been asked this many times throughout my short science policy career and ...importantly, how, of a science policy career.  Who does science policy? I’ve intentionally started with... many science policy professionals that entered the field from a more traditional policy background, learning...to start. What is science policy? THE IMPOSSIBLE QUESTION. Science policy is the work you do to creatively...pressures including economic, cultural, and political. Science policy looks differently depending on how and...

...Figure 1: EvolvR consists of an error-prone polymerase (PolI) that is fused to a nicking Cas9. A guide ...nicks its target DNA (left). Cas9 dissociates and PolI binds (middle) and extends from the 3' end of the...which was expected since the processivity window of PolI is 15-20 bp. EvolvR’s on-target mutagenesis rate...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...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...

...Plasmid tRNA Synthetase Expression Host pMAH-POLY Polyspecific aminoacyl-tRNA Synthetase Mammalian Cells ...protein of interest: you just don’t know it yet! The poly-histidine tag often used for protein purification...Green Tc: CCPGCC 50 40-72 Intact Cells Cy3NTAc Poly His  44 35-63 Plasma membrane protein, permeabilized...Lumio Red Tc: CCPGCC 48 38-70 Intact Cells Cy3NTAc Poly His  63 50-90 Plasma membrane protein, permeabilized...Lumio Red Tc: CCPGCC 54 42-78 Intact Cells Cy3NTAc Poly His  65 52-94 Plasma membrane protein, permeabilized...

...J, Plank C, Aneja MK, Rudolph C (2019) Segmented poly(A) tails significantly reduce recombination of plasmid...vectors (LTRs and ITRs), plasmids with multiple polyA sites, and other plasmids that contain repeating...

...transcription started by 5' LTR by the addition of a poly A tract just after the R sequence. This post... a structural precursor protein, and Pol is a polymerase. Envelope gene (may be pseudotyped to alter infectivity...Integrase components. Transfer cPPT Central polypurine tract; recognition site for proviral DNA synthesis...

...ends were modified slightly with either a 1-7 base poly(U)tail in tRNA-gRNA constructs or a 1-4 base tail...separated by 28 base Csy4 recognition sites from a polycistronic transcript. Although this method shortens construct...requires coexpression of Csy4 with Cas9 and the polycistronic transcript. As nucleases may have off-target... of gRNA cleavage.   Xie et. al constructed polycistronic glycine tRNA-gRNA genes (PTGs) in the forms ...Bacteria and viruses have illustrated the power of polycistronic gene organization, and its translation to CRISPR...

...secondary antibody depends on whether you are using poly- or monoclonal primary antibodies. Some secondary...question you may have is whether a monoclonal or polyclonal antibody is better for your experiment. Monoclonal...antibody clone and therefore a single isotype while polyclonal antibodies contain a mix of different isotypes...tend towards more reproducible results. Since polyclonal antibodies contain a variety of clones and thus...secondary antibodies instead (B).   Since a polyclonal primary antibody contains a mix of different ...reactive secondary antibodies when you’re using polyclonal primary antibodies.  I hope this post has explained...

...used for purification as elution requires low pH poly His HHHHHH  0.8 Affinity and Purification Very small.... Arguably the simplest affinity tag is the polyhistidine (His) tag. Small and unlikely to affect function...the tag size quickly – modifying the length a polyhistidine tract does not greatly alter the size of the...

...reading frames (ORFs) which often terminate with a poly(A). ORF2 encodes endonuclease and reverse transcriptase...They contain two genes: gag and pol. The pol polyprotein encodes the reverse transcriptase and integrase...

... 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 ...

...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 ...