...80 °C. Titer All titering is performed on lentiviral preparations that have been stored at -80 °C and ...of AAV preparations is assayed by comparing the relative stoichiometric ratios of the viral capsid proteins...SYPRO Ruby staining and the molecular weight and relative intensity of the viral capsid proteins are analyzed...Viral capsid proteins VP1, VP2, and VP3 are shown relative to the total protein present in the sample. Sterility...vectors consist of full particles (white arrowhead) relative to empty particles (black arrowhead). Scale bar...Preparations are then aliquoted and stored at -80 °C. Titer Titering is either performed by Addgene or ...

...Intuitively, one would expect that assembling relatively small, repeat-laden DNA regions into a single...Center (www.taleffectors.com) . Whether you work in a C. elegans lab and have been struggling to mutate a ..., Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic...using engineered TALENs. Sander JD, Cade L, Khayter C, Reyon D, Peterson RT, Joung JK, Yeh JR. Nat Biotechnol...at Massachusetts General Hospital also recently released a TALEN kit through Addgene. This kit, first described...

...stored at 4 °C for short-term storage. For long-term storage they should be stored at -20 °C in aliquots...colored product that accumulates over time which is relative to the amount of enzyme present in each well. ... categories: Quantification - used to quantify relative amounts of protein. Includes: Western blot ELISA... western blots or ELISAs, are used to quantify relative amounts of protein. Western blot Western blots...proteins separated by size on an SDS-PAGE gel. Though relative quantification can be done, western blots primarily...referred to as housekeeping genes). This allows for relative comparison of expression between different samples...conjugated to fluorophores should never be stored below 4 °C. Certain isotype subclasses, such as IgG3, aggregate...

...: 95 °C for 15 min, 35 cycles of (95 °C for 30 sec, 60 °C for 1 min, 72 °C for 1 min), and 72 °C for 10...following parameters: 37 °C for 30 min; 95 °C for 5 min and then ramp down to 25 °C at 5 °C/min. Dilute oligos...parameters: Cycles 1-20 (37 °C for 5 min, 20 °C for 5 min); Cycle 21 (80 °C for 20 min). These cycling ...Incubate at 30 - 37 °C for 24 - 72 hr. 30 °C may enhance genome editing efficiency, but 37 °C is acceptable....cells to incubate at 37 °C for 3 - 7 days and allow the clones to incubate at 37 °C for 7 - 14 days. Vary...thermocycler and run the following program: 65 °C for 6 min, 98 °C for 2 min to extract gDNA. Measure the DNA...clones. Run sample in thermocycler: 65 °C for 6 min and 98 °C for 2 min to extract gDNA. Screen each clone...

...to generate iPSCs. The generation of iPSCs is relatively simple in concept: ectopically express a cocktail... state using a cocktail of factors (Oct3/4, Sox2, c-Myc, and Klf4) that are known to maintain pluripotency...Doxycycline-inducible expression of human Oct4, Sox2, Klf4, and c-Myc from four separate lentiviral plasmids A drug-...for the expression of human Oct4, Klf4, Sox2, and c-Myc Human Induced Pluripotent Stem Cells Produced ...lentiviral vector for the expression of human Oct4, Klf4, c-Myc, and Sox2 Integrative Analyses of Human Reprogramming... Lentivirus Human Expression of human Klf4, Oct4, c-Myc, and Sox2 as VP-16 transcriptional activating ...polycistronic expression of human Oct4, Klf4, Sox2, c-Myc and hairpin RNA p53 for single plasmid reprogramming...

...window) C-terminal Anti-MECP2 (M6818) mouse antibody from Sigma (Link opens in a new window) C-terminal...diagnostic criteria, defined as the presence of relatively normal early development, regression with a loss...localization signal (not essential for localization) the C - t erminal D omain (CTD) The most common missense... than mutations further downstream in the NID and C-terminus. A large number of mutations are known to...26944080 (Link opens in a new window) Alysson Muotri c.806delG 806delG G269Afs*20 M Fibroblasts & iPSC Unpublished... of treatments and cures for Rett Syndrome and related MECP2 disorders. The RSRT has a comprehensive, ...new window) PMID: 10508514 Archer et al. 2007. Correlation between clinical severity in patients with Rett...

...researchers also use bacteria like E. coli and their relatively well understood biology to answer many interesting...construct, grown at the permissive temperature (32°C) to allow transposition, and the construct is then... by growing at the non-permissive temperature (42°C). Transposition is efficient enough that selection...TALEN Gang Bao Gateway cloning and expression of a C-terminally His-tagged TALEN fused to an E. coli codon...protein fusion: contains empty vectors with N and C-terminal fluorescent protein tags that can be used... pET15-MHL 26092 T7-lacO (lactose/IPTG inducible) C-terminal TEV cleavable 6x His Cheryl Arrowsmith Empty...Lactose/IPTG inducible) Various combinations of N and C-terminal Strep and His tags Thorben Dammeyer Lactose...

...3rd 4 78,637 Broad GPP Humagne Set C and Humagne Set D 172650 (Set C) 172651 (Set D) Knockout Human Doench... for genetic screening experiments, due to the relative ease of designing gRNAs and the ability to modify...gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources ...pneumoniae M. tuberculosis M. smegmatis Green monkey ( C. sabaeus ) Kaposi's Sarcoma-associated Herpes Virus...3rd 4 2,852 Bonifacino Lab Human ubiquitination-related proteins CRISPR KO library 174592 Knockout Human...

...CCL1 chemokine (C-C motif) ligand 1 I-309, P500, SCYA1, SISe, TCA3 CCL11 chemokine (C-C motif) ligand 11...chemokine (C-C motif) ligand 13 CKb10, MCP-4, MGC17134, NCC-1, NCC1, SCYA13, SCYL1 CCL14 chemokine (C-C motif...chemokine (C-C motif) ligand 19 CKb11, ELC, MGC34433, MIP-3b, MIP3B, SCYA19 CCL2 chemokine (C-C motif) ligand...chemokine (C-C motif) ligand 20 CKb4, LARC, MIP-3a, MIP3A, SCYA20, ST38 CCL21 chemokine (C-C motif) ligand...CCL24 chemokine (C-C motif) ligand 24 Ckb-6, MPIF-2, MPIF2, SCYA24 CCL25 chemokine (C-C motif) ligand 25...chemokine (C-C motif) ligand 26 IMAC, MGC126714, MIP-4a, MIP-4alpha, SCYA26, TSC-1 CCL27 chemokine (C-C motif...CCL28 chemokine (C-C motif) ligand 28 CCK1, MEC, MGC71902, SCYA28 CCL3 chemokine (C-C motif) ligand 3 ...

...transgenic nematodes, including C. elegans and C. briggsae. Other related miniMos plasmids can be found ... below lists plasmids that contain a worm ( C. elegans, C. briggsae , etc.) gene or sequence: ID Plasmid...Heritable genome editing in C. elegans via a CRISPR-Cas9 system. Cloning-free CRISPR for C. elegans , which uses...General Tools Fire Lab C. elegans Vector Kit - Andrew Fire Lab. A set of vectors for C. elegans research, ...distributes stocks of C. elegans. Silencing Genomes - Cold Spring Harbor's guide to RNAi in C. elegans WormAtlas...concerning the genetics, genomics and biology of C. elegans and related nematodes. WormBook - A comprehensive, ...peer-reviewed chapters covering topics related to the biology of C. elegans and other nematodes. Wormbuilder...

.... A., Amrani, N., Peng, C., Kramme, C., Savic, N., Pacesa, M., Rodríguez, T. C., Stan, T., Tysinger, E...Dalvai, M., Loehr, J., Jacquet, K., Huard, C. C., Roques, C., Herst, P., Côté, J., & Doyon, Y. (2015)....Heussler, G. E., Hearne, C. C., Qu, J., Inclan, Y. F., Hawkins, J. S., Lu, C. H. S., Silvis, M. R., Harden...this strategy yields C to A base edits, whereas in mammalian cells, it produces C to G edits. Adenine ... Cas3 must be paired with the Cascade ( C RISPR- as sociated c omplex for a ntiviral de fense) complex...and Cas12a (also known as Cpf1) Cascade C RISPR- as sociated c omplex for a ntiviral de fense; a complex..., S., Bietz, A., Waldhauer, M. C., Börner, K., Fakhiri, J., Schmelas, C., Dietz, L., Grimm, D., Correia...

...window) Poulin JF, Caronia G, Hofer C, Cui Q, Helm B, Ramakrishnan C, Chan CS, Dombeck DA, Deisseroth K...Navarro M, Burnham N, Cristiano C, Dorrier CE, Tipton GJ, Ramakrishnan C, Kozicz T, Deisseroth K, Thiele...Markovic M, Wolff SB, Ramakrishnan C, Fenno L, Deisseroth K, Herry C, Arber S, Lüthi A. 2016. Midbrain ... in order to enable three-feature cell targeting. C) Example of viral targeting of cells expressing Cre...processing, producing a functional molecular tool (C,F). Implementation The following resources may be ...Vcre No References Fenno LE, Mattis J, Ramakrishnan C, Hyun M, Lee SY, He M, Tucciarone J, Selimbeyoglu ...Grosenick L, Zalocusky KA, Bernstein H, Swanson H, Perry C, Diester I, Boyce FM, Bass CE, Neve R, Huang ZJ, Deisseroth...

... Backbones Flag Epitope tag c-Flag pcDNA3 or FLAG-HA-pcDNA3.1 - C-terminal Flag or N-terminal FLAG-HA...pNIC-CTHF - C-terminal TEV-His6-Flag for bacterial expression pFA6a vectors - PCR-based C-terminal...Epitope tag pcDNA3.1-HA or c-Flag pcDNA3 - Mammalian expression vector with N- or C-terminal HA tag pInducer20...trp mTagBFP2-pBAD - Protein expression vector with C-terminal mTagBFP2 tag pNIC-GST-TEV-GG - Cloning of...Gileadi lab Worm unc-54, variety of worm gene promoters C. elegans vector kit - Collection of...of plasmids from the Fire Lab L4440 - RNAi in C. elegans Also see our Worm Plasmids and...purification. Just remember to remove the stop codon for C-terminal tags and omit the start codon for N-terminal...

...NR1F3, RORG, RZR-GAMMA, RZRG, TOR RAR-related orphan receptor C RXRA Plasmids RXRA, NR2B1 retinoid X receptor... GCR, GR, GRL nuclear receptor subfamily 3, group C, member 1 (glucocorticoid receptor) NR3C2 Plasmids... MR, NR3C2VIT nuclear receptor subfamily 3, group C, member 2 NR4A1 Plasmids NR4A1, GFRP1, HMR, N10, NAK...estrogen-related receptor alpha ESRRB Plasmids ESRRB, DFNB35, ERR2, ERRb, ESRL2, NR3B2 estrogen-related receptor...Engineering Guide Empty-Backbone Resource Image is released to public domain by author. Source: Wikipedia,...Plasmids ESRRG, ERR3, ERRgamma, NR3B3 estrogen-related receptor gamma HNF4A Plasmids HNF4A, RP5-1013A22.1...MGC119326, MGC119329, RZR-ALPHA, DKFZp686M2414 RAR-related orphan receptor A RORB Plasmids RORB, NR1F2, ROR-BETA...

...pathway in vivo . DULIP ( DU al L uminescence-based C o- I mmunoprecipitation) plasmids : Luminescence-based...fusing NanoLuc® and Venus fluorophore to Troponin C Ca 2+ binding domain. Nano-lanterns , Enhanced Nano-lanterns...87075 pLenti6.2-ccdB-Nanoluc NanoLuc® Creation of C-terminal Nanoluc fusions using Gateway cloning. Lentival...Taipale 87065 pcDNA3.1-ccdB-Renilla Renilla Creation of C-terminal Renilla luciferase fusions using Gateway ...Taipale 87066 pcDNA3.1-ccdB-Firefly Firefly Creation of C-terminal Firefly luciferase fusions using Gateway ... 87074 pLenti6.2-ccdB-Firefly Firefly Creation of C-terminal Firefly luciferase fusions using Gateway ...Alberto Macho 174051 pGWB-cLUC Firefly Creation of C-terminal Fifrefly luciferase fragment for split-luciferase...

...Eisenhoffer 131781 pCS2+PhyBΔCreC CreLite - Red light-inducible Cre; C terminus Cre fused to PhyBdelta ... a circular piece of DNA (and is not maintained). C) If the sites are on separate DNA molecules (in trans... Cre: Cre recombinase is split in half into N and C terminal Cre fragments (NCre and CCre, respectively...Bacterial Ekker 47551 pDD104 (Peft-3::Cre) Cre eft-3 C. elegans Goldstein 48201 CAG-GFP-IRES-CRE Cre and ...system CAG Mammalian Pelczar 51268 pCAG-Co-InCreC C-terminal component of the Co-InCre system CAG Mammalian...GFP (CRE-DOG) EF-1 alpha AAV Cepko 69571 pAAV-EF1a-C-CreintG Cre recombinase dependent on GFP (CRE-DOG)...dependent on GFP (CRE-DOG) CAG Mammalian Cepko 69573 pCAG-C-CreintG Cre recombinase dependent on GFP (CRE-DOG)...

...for Tet transregulators. Krueger C, Pfleiderer K, Hillen W, Berens C. Biotechniques . 2004 Oct;37(4):546...developed. tTA was created by fusing tetR with the C-terminal domain of VP16 (virion protein 16), an essential...encoded tetracycline resistance. Hillen W and Berens C. Annu Rev Microbiol . 1994;48:345-69. PubMed . Transcriptional...cells. Yao F, Svensjö T, Winkler T, Lu M, Eriksson C, Eriksson E. Hum Gene Ther . 1998 Sep 1;9(13):1939...biomedical research. Bockamp E, Maringer M, Spangenberg C, Fees S, Fraser S, Eshkind L, Oesch F, Zabel B. Physiol...rTetR, which reversed the phenotype and created a reliance on the presence of tetracycline for induction,...

...System, AmpR 26100 pFHMSP-LIC-C N/A Expression vector for secreted proteins with C-terminal hexahistidine tag...dimensional structures of proteins of medical relevance, and place them in the public domain without restriction...TEV cleavage, Bac-to-Bac Expression System, AmpR Related Resources SGC: http://www.thesgc.org/ SGC Toronto...

...LRRK2 C-His LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40376 pBACgus/LRRK2 1867-2141 C-His...2161 C-His LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40378 pBACgus/LRRK2 1867-2176 C-His...2161 C-His_T2031E LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40381 pBACgus/LRRK2 C-His_G2019S...2161 C-His_G2019S LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40383 pBACgus/LRRK2 C-His_C2024S...mCherry-MAPTau-C-10 MAPT mCherry CMV Parkinson's, FTD Michael Davidson 55311 mTagBFP2-MAPTau-C-10 MAPT mTagBFP2... mKO2-MAPTau-C-10 MAPT mKO2 CMV Parkinson's, FTD Michael Davidson 58112 tdTomato-MAPTau-C-10 MAPT tdTomato...CMV ALS Alicia Pickrell 131792 pHAGE-TBK1-C-FLAG-HA TBK1 STOP codon CMV ALS Alicia Pickrell 131889 pDONR221...

...gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources ...which a reverse transcriptase (RT) is fused to the C terminus of Cas9 H840A nickase. The fusion enzyme ...transcriptase. Engineering of the PE enzyme and related components have helped to improve the efficiency...the reverse transcriptase. Many PE enzymes and related tools are available: PE2 - introduction of five...