...of bacterial genes. Molecular Systems Biology, 14(3):e7899. doi: 10.15252/msb.20177899. PMID: 29519933...attachment site of the phage 186. Detailed information about the repression system can be found in this article...29519933 . bioRxiv preprint . Detailed information about chromosomal integration of pLC97 can be found in this...

...). DeChenne-Peters et al. PLoS One. 2023 Mar 9;18(3):e0282170. PMID: 36893201 (Link opens in a new window...integrating MDH-related research into their teaching laboratories. They encourage instructors teaching different...research community and facilitates scientific collaborations between institutions. With these plasmids, ...regulation and post-translational modifications, and metabolic pathways and protein interactions. Plasmids A ...projects, find teaching resources and protocols, learn about regional hubs, access a teaching mentor, and more....2021.43 (Link opens in a new window) External Collaboration Results in Student Learning Gains and Positive...

...Transfer plasmids with sequence from the 5′ to the 3′ ITR exceeding 4.7 kb Transfer plasmids with a cargo... by email will supersede the general guarantees above. We will have our expert viral vector production...supersede the general estimated turnaround time above. After starting viral production, if we determine...

... consists of 5 individually packaged plasmids and 3 individually packaged strains, which are shipped as...Dove et al., Nature 1997 ) and later modified by Pabo and colleagues ( Joung et al., PNAS 2000 ). The ...

...cutting enzymes: NotI at the 5' end and AscI at the 3' end. These inserts can then be sub-cloned into a ...important in cellular recognition processes. Laboratories interested in these approaches have created ... and secreted proteins Mouse sperm Mouse egg Laboratories using AVEXIS: Christian Sollner, Max Planck ...

... work in your mouse line, see Supplementary Table 3 (Link opens in a new window) . Browse Available PHP.eB...Inhibitory - AAV-PHP.eB with GRE or transgenic animal L2/3 & L4 - AAV.CAP-B10 with GRE or transgenic animal Excitatory...systemic serotypes from the Viviana Gradinaru laboratory and the Caltech CLOVER Center (Link opens in ...AAV Decision Tree The AAV decision tree graphic above helps you choose the best capsid for your cell type...

...Combinatorial Strategies Connie Cepko 2:45 PM Break 3:00 PM (Group Discussion) Technologies and Resources...Learn about Addgene's 2018 AAV Education and Development (AAVED) workshop series. The... Institute Katherine Matho Cold Spring Harbor Laboratory Gowan Tervo Janelia Research Campus Sarada Viswanathan...Institute Katherine Matho, Cold Spring Harbor Laboratory Gowan Tervo and Sarada Viswanathan, Janelia Research...

... rpoZ- on plates lacking histidine and containing 3-AT Scot Wolfe Bacterial one-hybrid system for testing...promotes growth on plates lacking His and containing 3-AT. FLIParaF.Ec-250n 18855 N/A FRET between CFP and...in vitro ? These, in addition to many questions about bacteria themselves, can be answered using model...overexpressing or knocking down a gene can tell you much about its function and/or alter cellular activity in a...basic biology of a bacterial gene or redirecting metabolic pathways to produce a therapeutic compound, there...once you get it right, you can learn many things about your protein’s function, structure, and interactions...molecules, light, temperature, or even intracellular metabolite levels. Different plasmids and, often times, ...

...genetic model organism due to their rapid life cycle (3 days from egg to adult) and easy lab maintenance. ... approach has also been developed by the Fire laboratory and described in Efficient Marker-Free Recovery...

...Cre lac Bacterial Ekker 47551 pDD104 (Peft-3::Cre) Cre eft-3 C. elegans Goldstein 48201 CAG-GFP-IRES-CRE...deltaG)-Cre Cre CMV RABV Jessell 73717 pMLS328 Cre eft-3 C. elegans Jorgensen 75267 pAAV-CIBN-CreC photo-activatable...Vivo Electroporation. Proc Natl Acad Sci. Jan 16;104(3):1027-32. doi: 10.1073/pnas.0610155104. Epub 2007 ...Learn about the Cre-Lox system and how it can be used for genetic rearrangements and recombination. Use...

...Fishell 83898-AAV1 pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic...Fishell 83898-AAV9 pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic...Fishell 83898-AAVrg pAAV-mDlx-ChR2-mCherry-Fishell-3 ChR2-mCherry fusion expression in forebrain GABA-ergic...particles that are suitable for in vivo use. Learn more about our viral service ID Viral Prep Description PI 65417...well as by our trusted partner labs. Learn more about our antibodies ID Recombinant Antibody Reactivity...

...TALEs for the masses. Rusk N. Nat Methods 2011 Mar;8(3):197. Pubmed PMID: 21473016 . Move over ZFNs. Nat ...researchers who are learning to wield them. Learn more about Addgene’s TAL effector and TALEN kits here . References...

...standard consisting of ECFP and EYFP connected by 3 GGSGGS repeats pET28CLY4 Peptide linker standard consisting...constructed using the cpFRET kit from the Pertz laboratory. Empty Vectors Encoding Fluorescent Proteins ...

...shRNA cassettes into the unique SnaBI site in the 3´-LTR). See here for other versions of pULTRA. Moore... Read our Lentiviral Guide for more information about: Lentiviral components 2nd versus 3rd generation...systems Viral production Frequently asked questions about lentivirus use Glossary of commonly used lentiviral... or shRNA. See article for detailed information about cloning & additional plasmids. Aebischer and Trono...

...variant 3' NGAG SpCas9 VQR variant  3' NGAN or NGNG  xCas9 3' NG, GAA, or GAT SpCas9-NG 3' NG ...SpCas9 3' NGG SpCas9 D1135E variant 3' NGG (reduced NAG binding) SpCas9 VRER variant 3' NGCG...CJ) 3' NNNNRYAC  Neisseria meningitidis (NM) 3' NNNNGATT  Streptococcus thermophilus (ST) 3' NNAGAAW...target sequence contained within the guide RNA and a 3-base pair sequence known as the Protospacer Adjacent...must contain a PAM sequence immediately downstream (3’) of the site targeted by the guide RNA. In the absence...researchers to target many different genomic loci. 3. Cleave target DNA resulting in a double-strand break...mediated DNA damage is a DSB within the target DNA ~3-4 nucleotides upstream of the PAM sequence. Cas9 Species...

... and breathing, and often results in death within 3-5 years from symptom onset. Disease mechanisms are...Find out about the plasmid collections available at Addgene that may be useful for neurodegeneration ...Scientists are studying protein aggregation, learning about the blood brain barrier integrity, and using CRISPR...with support from the BRAIN Initiative. Jackson Laboratory (Link opens in a new window) A collection of ...

...assist with restriction enzyme digestion (usually 3-6bp) Restriction Site: Your chosen restriction site...Primer will use the sequence 5'-ATGTGGCATATCTCGAAGTAC-3' for the region that binds the ORF and we will add...our Forward Primer 5'-GAATTCATGTGGCATATCTCGAAGTAC-3'. Many restriction enzymes do not cut DNA efficiently...more information). Thus, we recommend that you add 3-6 bases upstream of your restriction site to improve... sequence of 5'-TAAGCAGAATTCATGTGGCATATCTCGAAGTAC-3'. For the Reverse Primer, the design is similar, but...including the stop codon (5'-TGGCATATCTCGAAGTACTGA-3'), then adding NotI (GCGGCCGC) and then TAAGCA to ...sequence of 5'-TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3' (30bp with 18bp of homology to the ORF). We now need...

...polymerase leaves a single deoxyadenosine (A) at the 3' ends of PCR products. The complimentary “T” in the...this enzyme recognizes the DNA sequence 5´-(C/T)CCTT-3' and digests double stranded DNA at this sequence.... is stored as a covalent bond between the cleaved 3′ DNA strand and a tyrosyl residue of topoisomerase... provide vectors or cloning arms with overhanging 3´ deoxythymidine (T) residues that are covalently linked...potential positive clones. Once you introduce your 3’-end “A” overhang insert, the magic of TOPO cloning...Reaction: Mix together the PCR product and TOPO Vector.  3. Incubate 5 Minutes at Room Temperature: You can place...proofreading polymerases will also remove all unpaired 3’ ends in your PCR product. If you need to decrease...

...ligase 25 °C 1 hour 3-base or 4-base T4 DNA ligase 25 °C 18 hours 3-base or 4-base T4 ...ligase 37 °C 1 hour 3-base or 4-base T4 DNA ligase 37 °C 18 hours 3-base or 4-base Bbsl-HF...-16 °C cycling 3-base Sapl, 1x NEBridge Ligase MM 37-16 °C cycling 3-base Table 1: Enzyme... the same overhang twice; (2) avoid palindromes; (3) no overhangs with the same three nucleotides in a...could be done using overhang sets that broke rules 3—5. Fidelity, it turns out, might depend more on the...must be used. Each of the tools works with either 3-base or 4-base overhangs, and with a variety of different...BspQI, 1x NEBridge Ligase MM 42-16 °C cycling 3-base  PaqCl®, 1x T4 DNA Ligase Buffer 37-16 ...

...Figure 3: BaF3 cells were transduced with H2B-FT virus and grown in varying concentrations of IL-3. Cells..., which depend on IL-3 to proliferate. The cells cultured with a low dose of IL-3 showed reduced proliferation...high IL-3 showed mostly blue fluorescence, indicating a quick cell cycle. Cells that had IL-3 withdrawn...Cells grown in high concentrations of IL-3 show a high proportion of blue fluorescence, indicating cell ...proliferation. BaF3 cells maintained in media without IL-3 show only red fluorescence. Image from Eastman et ...Streptococcus canis Cas9 nuclease that has a broad 5′-NNG-3′ PAM. Get the plasmids at Addgene The Human GlycoGene... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's...