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Escherichia coli strain xl10 gold

Manufactured by Agilent Technologies
Sourced in United States

Escherichia coli strain XL10-Gold is a competent bacterial strain commonly used in molecular biology applications. The strain is engineered to improve transformation efficiency and enable the propagation of plasmid DNA.

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4 protocols using escherichia coli strain xl10 gold

1

Plasmid Construction and Cloning

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Construction of the plasmids is described in detail in Supplementary Tables S1 and S2. DNA sequences for AmDODA and CcTyr can be found in DNA sequences S1 and S2. Escherichia coli strain XL10-Gold (Agilent Technologies) was used for cloning.
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2

Preparation and Use of HIV-1 Env Plasmids

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Escherichia coli strain XL-10 gold (Agilent) and Stbl2 cells (Invitrogen) were products of Novagen Inc. (Madison, WI). Thermostable DNA polymerase (PfuUltra) was obtained from Stratagene Inc. (La Jolla, CA). Custom-oligonucleotide primers were supplied by Integrated DNA Technologies (IDT). DNA plasmids encoding HIV-1BaL Env (catalog no. 11445) and NL4-3 R E Luc+ were obtained from the NIH AIDS Reagent Program, Division of AIDS, NIAID, and were a kind gift of Dr. J Mascola and Dr. N Landau, respectively. All other reagents used were of the highest analytical grade available.
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3

Growth Characterization of Streptococcus pneumoniae

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Streptococcus pneumoniae TIGR4 (accession: AE005672.3) wild-type strain (WT) and its derivative strains were cultured in Todd-Hewitt broth (BD Biosciences) supplemented with 0.2% yeast extract (BD Biosciences) (THY) with or without antibiotics at 37°C. For growth measurements, overnight cultures of each strain were back-diluted 1:50 into fresh THY and grown at 37°C. Growth was monitored by measuring optical density at 600 nm (OD600) every 0.5–1 hour.
Escherichia coli strain XL10-Gold (Agilent Technologies) was used as a host for derivatives of the pDCerm plasmid containing the erythromycin-resistance cassette [48]. E. coli strains were cultured in Luria-Bertani broth (LB) (Nacalai) at 37°C with agitation. For selection and maintenance of recombinant strains, antibiotics were added to the medium at the following concentrations: erythromycin (Sigma-Aldrich), 400 μg/mL for E. coli and 5 μg/mL for S. pneumoniae; spectinomycin (Wako), 120 μg/mL for S. pneumoniae.
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4

Streptococcus pyogenes Culturing and Manipulation

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Streptococcus pyogenes M1T1 strain 5448 (accession: CP008776.1) was isolated from a patient with toxic shock syndrome and necrotizing fasciitis that is genetically representative of a globally disseminated clone associated with invasive S. pyogenes infections (Kansal et al., 2000 (link)). S. pyogenes strains were grown at 37°C in a screw-cap glass tube (Pyrex; Iwaki Glass, Tokyo, Japan) filled with Todd-Hewitt broth (BD Biosciences, San Jose, CA, USA) supplemented with 0.2% yeast extract (BD Bioscience) (THY broth) in an ambient atmosphere and standing cultures. To obtain cultures for experiments and observe pH change, overnight cultures of S.pyogenes were back diluted 1:50 into fresh THY broth or phenol red broth (Sigma Aldrich, St Louis, MO, USA) supplemented with 30 mM arginine. CFUs were determined by plating diluted samples on THY blood agar.
Escherichia coli strain XL-10 Gold (Agilent Technologies, Santa Clara, CA, USA) was used as a host for derivatives of plasmids pSET4s (Takamatsu et al., 2001 (link)) and pQE30 (Qiagen, Hilden, Germany). E. coli strains were cultured in Luria-Bertani medium (Nacalai Tesque, Kyoto, Japan) at 37°C with agitation. For selection and maintenance of strains, antibiotics were added to the medium at the following concentrations: spectinomycin, 100 μg/mL for S. pyogenes and E. coli: carbenicillin, 100 μg/mL for E. coli.
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