...circular assemblies. While Golden Gate Assembly worked for all the assembly types, circular assemblies yielded... circular assembly of the T7 genome had lower efficiency than the 10-piece circular assembly, with the...accomplished with Golden Gate Assembly—even when getting help from DAD. The 35-piece assembly mentioned above had...limit for Golden Gate Assembly size. Ultimately, data-driven Golden Gate Assembly greatly increases the...complex Golden Gate Assembly, a great place to start is the 24-piece lac operon assembly deposited by NEB...-pot Golden Gate assemblies of unprecedented complexity using data-optimized assembly design. PLOS ONE...If you’ve ever used Golden Gate Assembly for cloning, you might be familiar with the rules of thumb for...

...insight to the now 10 member board at Addgene. Let’s meet the newest board members and get to know them...looking for new board members. Paratore: Kristen Darby, who’s also a board member, reached out and asked...Dorr and Crowell & Morning. Karen was a founding member of Mass Medical Angels, a venture mentor at MIT...certified health insurance executive (CHIE). He is a member of both the Internal Institute of Auditors and ...blog Resources on Addgene.org Meet the other members of Addgene's Board of Directors Find careers at...

...basics of Gibson assembly as a parody of “Breakfast at Tiffany’s.” Overview of Gibson assembly The Gibson assembly...gain in popularity. Gibson assembly is well known for allowing easy assembly of multiple linear DNA fragments...Miller Lab Protocol). The Gibson assembly process can be used to assemble up to 6 fragments in one step,...design primers for Gibson Assembly. For a simple example of using Gibson assembly, imagine that you want ...annealing and priming of neighboring assembly fragments. Gibson assembly is commonly used in synthetic biology...in 2013 found that Gibson assembly was one of the most commonly used assembly methods (Kahl 2013). However...cut out via CRISPR and then assembled into a vector using Gibson assembly. In this case the vector was...

...to different lipid compositions of C. elegans membranes, which might beneficially influence the rhodopsin...

...podcast segment A fundamental approach to studying membranes, organelles, and other subcellular compartments...

...started, the hardest part of the process was to embed personality in pieces of plastic, but it’s gotten...pictures, one for each position in a movement, and assembling the many pictures together to form the full movement...when we started was creating a LEGO stage that resembles a real laboratory. It only took one night to build...

... different barcodes to each of the gRNAs and to embed these barcodes within the gRNA sequences. This allowed...

...fluorescent voltage indicator. Archon1 can embed itself into the cell membrane where cell voltage can then be measured...

...subfamily C, member 3 NKG2-E, NKG2E KLRC4 killer cell lectin-like receptor subfamily C, member 4 FLJ17759...subfamily D, member 1 CD94 LGMN legumain AEP, LGMN1, PRSC1 LTA lymphotoxin alpha (TNF superfamily, member 1) LT...family A, member 2 PDA2, PDI, PDIP, PDIR PDIA3 protein disulfide isomerase family A, member 3 ER60, ERp57...FLJ12287, RP35, SEMAB, SEMB SEMA4B sema domain, immunoglobulin domain (Ig), transmembrane domain (TM) and short...MARVEL transmembrane domain containing 2 CKLFSF2, MGC39436 CMTM3 CKLF-like MARVEL transmembrane domain...CKLF-like MARVEL transmembrane domain containing 4 CKLFSF4 CMTM5 CKLF-like MARVEL transmembrane domain containing...MARVEL transmembrane domain containing 7 CKLFSF7, FLJ30992 CMTM8 CKLF-like MARVEL transmembrane domain...

... D, member 1 NR1D2 Plasmids NR1D2, RVR, BD73, EAR-1R nuclear receptor subfamily 1, group D, member 2 NR1H3...Browse the nuclear receptor plasmid collection assembled in collaboration with the Nuclear Receptor Signaling... domain by author. Source: Wikipedia, accessed November 6, 2013. Author: Boghog2 Nuclear receptors are...research labs from around the globe, Addgene has assembled a Nuclear Receptor plasmid collection for the ...NROB1, SRXY2 nuclear receptor subfamily 0, group B, member 1 NR1D1 Plasmids NR1D1, EAR1, hRev, THRA1, THRAL...LXRA, RLD-1 nuclear receptor subfamily 1, group H, member 3 NR2E1 Plasmids NR2E1, RP3-429G5.1, TLL, TLX, ..., XTLL nuclear receptor subfamily 2, group E, member 1 NR2F1 plasmids NR2F1, COUP-TFI, EAR-3, EAR3, ERBAL3...

...PI FusX TALEN Assembly System Contains 336 pFusX clones allowing for Golden Gate assembly of 15-16 RVD ...expression. Feng Zhang LIC TAL Effector Assembly Kit Assembly by ligation-independent cloning (LIC). Validated...plasmids of pre-assembled trimer and tetramer RVD domains designed for rapid TALEN assembly. Kiran Musunuru...expression of Voytas Golden Gate-assembled TALEN, both in microinjected embryos and transfected cells. TALEN...accompanying documentation allow one to efficiently assemble TALEN constructs with custom repeat arrays. TALEN...Bogdanove Joung Lab TAL Effector Engineering Reagents Assembly via serial ligation. Validated in zebrafish somatic...Ekker Platinum Gate TALEN Kit Platinum TALENs assembled by Golden Gate. Validated in several species. ...

...Parkinson's Michael J Fox Foundation MJFF 40427 pEMB017 NP_9409803_LRRK2_1867_2161_L1914N_V2048T_F2083Y...Parkinson's Michael J Fox Foundation MJFF 40499 pEMB018 / LRRK2_BC_1300_2136 LRRK2 His polH Parkinson's...'s Michael J Fox Foundation MJFF 40500 pEMB018 / LRRK2_BC_1300_2148 LRRK2 His polH Parkinson's Michael...Michael J Fox Foundation MJFF 40501 pEMB018 / LRRK2_BC_1300_2170 LRRK2 His polH Parkinson's Michael J Fox Foundation... Foundation MJFF 40502 pEMB018 / LRRK2_BC_1300_2210 LRRK2 His polH Parkinson's Michael J Fox Foundation...Foundation MJFF 40503 pEMB018 / LRRK2_BC_1309_2136 LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40504... 40504 pEMB018 / LRRK2_BC_1309_2170 LRRK2 His polH Parkinson's Michael J Fox Foundation MJFF 40505 pEMB018...

...37351 pQC membrane TdTomato IX Membrane Palmitoylation TdTomato Connie Cepko 22479 FUmGW Membrane Palmitoylation...pCAG-mGFP Membrane Palmitoylation sequence from GAP43 EGFP Connie Cepko 61099 Lck-GFP Membrane Palmitoylation...Mitochondria-Outer Membrane TOMM20 mCherry* Michael Davidson 118736 pBOB-CARMIL2 BH domain-GFP Plasma Membrane CARMIL2...protein of interest with known markers. Addgene has assembled a collection of fluorescent fusion proteins that... Michael Davidson 98822 Lck-mTurquoise2 Plasma Membrane Lck N-terminal lipidation signal mTurquoise2 Dorus... Dorus Gadella 98821 Lck-mScarlet-I Plasma Membrane Lck N-terminal lipidation signal mScarlet-I Dorus ... Vamsi Mootha 49153 GFP-Mff Mitochondria-Outer Membrane Mff acGFP1 Gia Voeltz 158007 pCMV-mGold-Mito-N...

...And Ligation) assembly method (Sander et al., Nat Biotechnol. 2011), the REAL-Fast assembly method (Reyon...or REAL-Fast assembly process or lists of plasmid ID numbers required for FLASH assembly. Links to additional... TAL effectors, including designed TALENs, and assembly methods for REAL, REAL-Fast and FLASH Genome...Ligation-based Automatable Solid-phase High-throughput) assembly method (Reyon and Tsai et al., Nat Biotechnol ...Biotechnol 2012). All three of these platforms assemble TAL effector repeat arrays on a single architectural framework...expression plasmids using REAL, REAL-Fast or FLASH assembly. This new version of ZiFiT Targeter enables users...these reagents are provided below: Joung Lab REAL Assembly TALEN Kit Individual REAL TALE Repeat Plasmids...

...separate vectors and then assembled into one construct using Golden Gate assembly. Gersbach MuLE System Mammalian...expression cassettes can be assembled using Golden Gate cloning or the Gibson Assembly. Chen Boosting CRISPR...this Golden Gate and Gateway based assembly method: plasmids for assembly of single gRNA vectors with various...Expression System Description PI Multiplex CRISPR/Cas9 Assembly System Kit Mammalian A system for constructing...-gRNA genes (PTGs) and then PTG cassettes are assembled into a Cas9 expressing vector. Yang A Robust CRISPR...multiple sgRNA expression cassettes, which can be assembled into the binary CRISPR/Cas9 vectors in one round...round of cloning by Golden Gate ligation or Gibson Assembly Liu A CRISPR/Cas9 toolbox for multiplexed plant...

...trafficking to the plasma membrane Acute disruption of the synaptic vesicle membrane protein synaptotagmin...measurements of membrane charges A FRET sensor enables quantitative measurements of membrane charges in live...Calcium Calcium sensor in astrocytes (GCaMP6f), membrane-tethered Khakh Lab pZac2.1 gfaABC1D-lck-GCaMP6f...2018 Sep;21(9):1272-1280. Lin Tian Calcium Plasma membrane targeted GCaMP6s calcium sensor A polarized Ca...+) Dynamics in the Mitochondrial Matrix and Intermembrane Space with the GZnP2 Sensor. ACS Chem Biol. ...sensor iATPSnFR1.0 and iATPSnFR1.1 (cytoplasmic, membrane-displayed, expression in neurons, astrocytes) ...reduced ph interference demonstrates augmented transmembrane chloride movement by gerbil prestin (SLC26a5...

...Finger Consortium Originally published in Addgene's December 2011 Newsletter Overview of TALEN Technology Imagine...can be made. Intuitively, one would expect that assembling relatively small, repeat-laden DNA regions into...from the University of Minnesota, arrays can be assembled in a matter of days. In an open access Nucleic...recently developed Golden Gate cloning method to assemble “multiple DNA fragments in an ordered fashion ...custom arrays consisting of 12-31 repeats can be assembled and inserted into a variety of backbones in just... in August, uses a serial ligation protocol to assemble arrays. Dr. Joung, co-founder of the Zinc Finger...early in 2011 and they described their kit for assembling TAL effector arrays in a 2012 issue of Nature...

...pluripotency during embryonic development. This “reprogramming” of the cell to an embryonic state holds immense...cell lineages. Pluripotent stem cells, such as embryonic cells, can potentially give rise to any other ...limited to related cell lineages. Pluripotent embryonic and multipotent adult stem cells have been studied...removes the possible ethical concerns related to embryonic stem cell use and has the major advantage that...Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors...Mouse Direct Reprogramming of Fibroblasts into Embryonic Sertoli-like Cells by Defined Factors. Cell Stem...via microRNA-dependent and -independent pathways. EMBO Mol Med. 2017 Aug;9(8):1117-1131. Parmar Fibroblasts...