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  1. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...containing T7 RNAP (DE3) F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) General protein expression BL21 (DE3...origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysS (CamR) Expression of toxic proteins...origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysE (CamR) Expression of toxic proteins...transcript half-life F- ompT lon hsdSB (rB- mB-) gal dcm rne131 (DE3) General expression; not recommended...required for expression F- ompT lon hsdSB (rB- mB-) gal dcm araB::T7RNAP-tetA General protein expression...repetative sequences.  F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) Δ(srl-recA)306::Tn10 (TetR) Expression...control of expression F- ompT lon hsdSB (rB- mB-) gal dcm lacY1 (DE3) Expression of toxic or insoluble...

  2. Inside Addgene's 10 Year Anniversary Gala

    Type
    Blog Post
    ...Chen. Last week, the Addgene 10-year Anniversary Gala celebrated all of the people who have helped Addgene...even posed for pictures with some Addgenies. As the gala came to an end, it was clear that the Addgenies ...journey has been a tough but memorable one. At the gala, one thing was clear: the sense of family is still...lovely play-by-play of Addgene's 10 Year Anniversary Gala. The Addgene lab often works as a very well-oiled...

  3. Plasmids 101: Blue-white Screening

    Type
    Blog Post
    ...expressed, the β-galactosidase enzyme can break down a dye-linked substrate called x-gal (5-bromo-4-chloro.... On the other hand, intact β-galactosidase produces pigment from x-gal (included in the transformation...enough time for any intact β-galactosidase to be expressed and process x-gal into blue pigment (16-20 hours... should be blue, indicating that your IPTG and x-gal are working as they should be. Don't rush the process...white colonies. Take care in making your plates: X-gal is light and temperature sensitive and needs to be...chloro-3-indolyl-β-D-galacto-pyranoside) into galactose and an insoluble blue pigment (4-chloro-3-brom-indigo...gene called lacZ that encodes for the enzyme β-galactosidase. Expression of the lac operon is induced by ...

  4. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    ...-type specificity! The Gal4/UAS system At a more detailed level, the Gal4/UAS system is a transcription...the UAS sequence are only expressed when Gal4 is expressed. Gal4 expression can be regulated by any chosen...Controlling multiple genes with Gal4/UAS system One can also use Gal4 to drive expression of multiple ...Applications of the Gal4/UAS system There are a number of fancy ways to use the Gal4/UAS system, and you... my recent paper to see the Gal4/UAS system in action (5). I use the Gal4/UAS system to knock down gene...down. To do this, Drosophila geneticists use the Gal4/UAS system. This incredibly useful, yet simple system...geneticist’s main workhorse to turn genes on or off. Gal4 is a transcriptional activator that binds to UAS...

  5. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    ...researcher, when the substrate X-gal is broken down by 𝛽-galactosidase, it turns into an insoluble blue...disrupting 𝛽-galactosidase activity. These cells, presumably the cell you want, cannot break down X-gal and are...transformation on ampicillin plates containing X-gal allows you to identify cells that took up the vector...on the lacZ gene. lacZ encodes the enzyme 𝛽-galactosidase which can hydrolyse lactose. Fortunately, for... is deleted, it creates a non-functional 𝛽-galactosidase. Complementing this mutation by expressing this...example on a plasmid, creates a functional 𝛽-galactosidase. Blue-white screens rely on vectors where the...

  6. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...featuring a guest blog post from Gal Haimovich of greenfluorescentblog. Gal will share his list of 10 things...experiments. In the meantime, if you haven't checked out Gal's blog, you should! He's got great articles and insight...

  7. Synthetic Biology - Algal

    Type
    Collection
    ...Plasmid Collections Synthetic Biology Algal Synthetic Biology: Algal SynBio Resources SynBio Home Browse...of synthetic biology plasmids for use in algae. Algal Plasmids Search the table by keyword or sort by ...

  8. Synthetic Biology - Fungal

    Type
    Collection
    ...Plasmid Collections Synthetic Biology Fungal Synthetic Biology: Fungal SynBio Resources SynBio Home Browse...of synthetic biology plasmids for use in fungus. Fungal Plasmids Search the table by keyword or sort by...

  9. Neurodegeneration Plasmid Collection

    Type
    Collection
    ...TARDBP GAL ALS Aaron Gitler 27465 pDuet TDP-43 Q331K TARDBP GAL ALS Aaron Gitler 27466 pRS426 Gal TDP43...TDP43 TARDBP GAL ALS Aaron Gitler 27467 pRS426 Gal TDP43 GFP TARDBP GFP GAL ALS Aaron Gitler 27468 pRS303...1185 p416 25Q GAL HTT GFP GAL1 Huntington's Susan Lindquist 1186 p416 103Q GAL HTT GFP GAL1 Huntington's...1187 p426 25Q GAL HTT GFP GAL1 Huntington's Susan Lindquist 1188 p426 103Q GAL HTT GFP GAL1 Huntington's...27458 pRS416 Gal TDP43 WT TARDBP GAL1 ALS Aaron Gitler 27459 pRS416 Gal TDP43 G294A TARDBP GAL1 ALS Aaron... pRS416 Gal TDP43 M337V TARDBP GAL1 ALS Aaron Gitler 27461 pRS416 Gal TDP43 Q331K TARDBP GAL1 ALS Aaron...pAG416-Gal-PR50 C9orf72 FLAG, Myc GAL1 ALS Aaron Gitler 84902 pAG416-Gal-PA50 C9orf72 FLAG, Myc GAL1 ALS ...

  10. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...  Plasmids for cGAL and split cGAL systems The Sternberg lab in 2016 created cGAL, a GAL-4 based bipartite...this system a promoter controls the expression of a GAL-4 driver that can bind to activation sequences upstream... 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust GAL4–UAS system for Caenorhabditis elegans...2018, the lab expanded on this tool by splitting cGAL4 in two and binding each half to a gp41-1-N-intein...split system, only cells that have both halves of cGAL can drive expression of your gene of interest.  ...Central PMCID: PMC5693259.  Wang, Han, et al. "Split cGAL, an intersectional strategy using a split intein...

  11. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...This post was contributed by Gal Haimovich of greenfluorescentblog. Be honest. Do you really know how...greenfluorescentblog. Thank You to Our Guest Blogger! Gal Haimovich, PhD, is a research fellow in the lab of...

  12. Adenovirus Plasmids

    Type
    Collection
    ...Vogelstein 16407 pAdEasy 2-GFP beta-gal Shuttle Test plasmid that contains β-gal and GFP; contains ∼10Kb genomic...

  13. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...resistance gal Mutation in galactose metabolism pathway Cells cannot grow on galactose only gyrA, .... F' mcrA Δ(mcrBC-hsdRMS-mrr) recA1 endA1 gyrA96 gal-thi-1 supE44 λ-relA1 Δ(lac-proAB)/F' proAB+lacIqZΔM15...Φ80lacZΔM15 ΔlacX74 recA1 araΔ139 Δ(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG fhuA::IS2 DB3.1 Streptomycin...screening, leucine auxotroph. F- endA1 recA1 galE15 galK16 nupG rpsL ΔlacX74 Φ80lacZΔM15 araD139 Δ(ara,...plasmid after preparation. rpsL thr leu thi lacY galK galT ara tonA tsx dam dcm glnV44 Δ(lac-proAB) e14- ...- Δ(ara-leu)7697 [araD139]B/r Δ(codB-lacI)3 galK16 galE15 λ- e14- mcrA0 relA1 rpsL150(strR) spoT1 mcrB1...(TetR) ∆(ara-leu) 7697 araD139 fhuA ∆lacX74 galK16 galE15 e14-  Φ80dlacZ∆M15 recA1 relA1 endA1 nupG rpsL...

  14. Plasmids 101: Protein Expression

    Type
    Blog Post
    ...proteins under the control of the galactose inducible promoter (GAL). Other commonly used promoters include...

  15. The Pleiades Promoter Project

    Type
    Collection
    ...Ple80 GABRA6 pEMS1434 EGFP/NLS Ple81 GAL pEMS1177 EGFP/NLS Ple84 GAL pEMS1180 EGFP/NLS Ple85 GCHFR pEMS1122...

  18. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ... et al. transduced rat neurons in vivo with beta-gal and other reporters, and they showed that expression...cells by a lentiviral vector. Naldini L, Blömer U, Gallay P, Ory D, Mulligan R, Gage FH, Verma IM, Trono ... Cartier N, Aubourg P, Fischer A, Cornetta K, Galacteros F, Beuzard Y, Gluckman E, Bushman F, Hacein-Bey-Abina...

  19. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...and GAL 80. TEF1 General expression mRNA Yeast transcription elongation factor promoter Constitutive  ...containing Gal4 binding sites, commonly used in Drosophila Specific Requires the presence of Gal4 gene to...with galactose; repressible with glucose  Can be used independently or together. Regulated by GAL4 and ...mammalian expression promoter from the human cytomegalovirus Constitutive  May contain an enhancer region...expression. Modulated by calcium and calmodulin. GAL1, 10 General expression mRNA Yeast adjacent, divergently...

  20. Centrifugation

    Type
    Protocol
    ...in a liquid sample. It achieves this by using centrifugal force, which is generated by spinning the sample...Pro-Tip Spin speed is often given as Relative Centrifugal Force (RCF) or Revolutions Per Minute (RPM), ...
Showing: 1 - 20 of 140 results