...His, TEV polH Parkinson's Michael J Fox Foundation MJFF 40375 pBACgus/LRRK2 C-His LRRK2 His polH Parkinson's...His polH Parkinson's Michael J Fox Foundation MJFF 40379 pBACgus/LRRK2 C-His_T2031E LRRK2 His polH Parkinson's...His polH Parkinson's Michael J Fox Foundation MJFF 40381 pBACgus/LRRK2 C-His_G2019S LRRK2 His polH Parkinson's... LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40385 pBACgus4x/Parkin PRKN polH Parkinson's...GST polH Parkinson's Michael J Fox Foundation MJFF 40407 LRRK2_1867_2143 LRRK2 His, TEV, GST polH Parkinson's...GST polH Parkinson's Michael J Fox Foundation MJFF 40409 LRRK2_1867_2134 LRRK2 His, TEV, GST polH Parkinson's... polH Parkinson's Michael J Fox Foundation MJFF 40505 pEMB018 / LRRK2_BC_1309_2187 LRRK2 His polH Parkinson's...

...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...

...lower background signals than polyclonal antibodies. However, polyclonal antibodies are more sensitive...clone that are specific to a certain epitope, or polyclonal, meaning there are many clones that are specific... antibodies are typically more expensive than polyclonal antibodies. Environmental factors, binding partners...available epitope on your protein of interest. Polyclonal antibodies are typically collected directly from...targeting various epitopes on the same antigen. Polyclonal antibodies are often more environmentally stable...protein in any form it may be present in. However, polyclonals can vary significantly from lot to lot, as immune...consider the following questions: Do you need polyclonal or monoclonal antibodies? Will you be using a...

...dependent 1, 5 Yizhar 154948 CaMKII-somBiPOLES-mCerulean CamKII BiPOLES (soma-targeted) mCerulean Constitutive... 5, 9, rg* Wiegert 154951 hSyn-DIO-somBiPOLES-mCerulean Syn BiPOLES (soma-targeted) mCerulean Cre dependent... Jaws stGtACR iC++ OPN3 PdCO PPO Bidirectional BiPOLES Promoter CAG CaMKII CBA Dlx EF1a/nEF Synapsin E2...

... PCR and surveyor. I used DNA polymerase Takara Ex Taq ™ DNA Polymerase for my genomic PCR, but couldn't...In our hands, Herculase II Fusion polymerase or Kapa Hifi Polymerase work very well. Some groups have ...your HR by doing Restriction Fragment Length Polymorphism (RFLP) ( see Figure 4 of the Cong, et. al, 2013...

...Marchantia polymorpha MpU6 none S. pyogenes Hara-Nishimura pMpGE_En03 71535 Other/Marchantia polymorpha MpU6...pyogenes Ventura pCRISPRyl 70007 Other/Yarrowia lipolytica SCR1'-tRNA yes, cut S. pyogenes LEU2 Wheeldon...pyogenes Draetta pCAS1yl 73226 Other/Yarrowia lipolytica yes, cut S. pyogenes LEU2 Yang pLCKO 73311 Mammalian...Gateway recombination-based cloning of complex polycistronic lentiviruses. Can express Cas9 with up to 3 ...in a single vector. sgRNAs are expressed as polycistronic glycine tRNA-gRNA genes (PTGs) and then PTG ...

...Ligation Buffer, 6.5 μl ddH 2 O, and 0.5 μl T4 Polynucleotide Kinase (PNK) (10,000 U/ml). NOTE: Phosphorylated...acceptable. The bulk sorted cells are composed of a polyclonal population exposed to sgRNA-A and sgRNA-B (see... band” in separate reactions. NOTE: Numerous polymerases may be used for step 6.3 . Run samples in a thermocycler...a single reaction. Optimize multiplexing in a polyclonal population ( i.e., bulk sorted cells) before ...biallelic deletion clones with a proofreading polymerase and clone the amplicons ( e.g., with a PCR cloning...

...mEGFP EZH2 Polycomb repressive complex 2 164500 POLR2A-mEGFP AICSDP-117 mEGFP RPB1 RNA polymerase II 168798...

...Mammalian TAA Simon Elsaesser 157925 pMAH-POLY-eRF1(E55D) POLY-eRF1(E55D) E. coli para-substituted phenylalanine...proteins for various purposes from therapeutics to biopolymers. Tips for Success Before beginning to reprogram...azidonorleucine (Anl) Mammalian ATG David Tirrell 64915 pMAH-POLY tyrosyl-tRNA synthetase E. coli pBof Mammalian TAG...

...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...

...      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...

...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...

...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...

... 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...

... 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 ...

...signaling, and transcription factor interactions (Kerppola, 2006). One of the first BiFC studies used the...construction, cellular expression, and quantification (Kerppola, 2006, 2013). These are described in more detail...RPACKIPNDLKQKVMNH have been used successfully (Kerppola, 2006). Flexible GS linkers may also work. You...analysis can be used in lieu of negative controls (Kerppola, 2006). Here, one of the endogenous PIPs is expressed...troubleshooting tips can be found in protocols by Kerppola (2006, 2013). BiFC vs. FRET: When to choose one...doi.org/10.7554/elife.38853 Hu C-D, Chinenov Y, Kerppola TK (2002) Visualization of Interactions among ... https://doi.org/10.1016/s1097-2765(02)00496-3 Kerppola TK (2006) Design and implementation of bimolecular...