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Difco amino acids
Amino Acids, supplied by Difco, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 92 stars, based on 6 article reviews
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amino acids - by Bioz Stars, 2021-01
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Transformation Assay:

Article Title: Identification of the Putative Protein Phosphatase Gene PTC1 as a Virulence-Related Gene Using a Silkworm Model of Candida albicans Infection ▿ Infection ▿ †
Article Snippet: .. SD-URA medium (synthetic dextrose medium without uracil) containing 0.67% yeast nitrogen base (YNB) without amino acids (Difco, Detroit, MI), 2% dextrose, and 0.072% CSM-URA (complete synthetic mixture without uracil) (QBiogene, Irvine, CA) was used for C. albicans transformation. .. For examination of C. albicans morphology, yeast or hyphal cells were produced by manipulation of the medium composition and temperature as follows.

Selection:

Article Title: The ZIP Transporter Family Member OsZIP9 Contributes To Root Zinc Uptake in Rice under Zinc-Limited Conditions 1The ZIP Transporter Family Member OsZIP9 Contributes To Root Zinc Uptake in Rice under Zinc-Limited Conditions 1 [OPEN]
Article Snippet: .. In a time-course experiment, ZHY3 expressing OsZIP9 or empty vector was grown in the Sc(-Uracil) medium containing 0.67% (w/v) yeast nitrogen base without amino acids (Difco), 2% (w/v) Glc, 0.2% (w/v) appropriate amino acid, and 2% (w/v) agar at pH 6.0 for selection. .. The yeast cells were first incubated in Sc(-Uracil) liquid medium with 50 m m of MES containing 2% (w/v) Gal or 2% (w/v) Glc (as a negative control) for 2 h, followed by washing three times with sterilized Milli-Q water (EMD Millipore).

Expressing:

Article Title: The ZIP Transporter Family Member OsZIP9 Contributes To Root Zinc Uptake in Rice under Zinc-Limited Conditions 1The ZIP Transporter Family Member OsZIP9 Contributes To Root Zinc Uptake in Rice under Zinc-Limited Conditions 1 [OPEN]
Article Snippet: .. In a time-course experiment, ZHY3 expressing OsZIP9 or empty vector was grown in the Sc(-Uracil) medium containing 0.67% (w/v) yeast nitrogen base without amino acids (Difco), 2% (w/v) Glc, 0.2% (w/v) appropriate amino acid, and 2% (w/v) agar at pH 6.0 for selection. .. The yeast cells were first incubated in Sc(-Uracil) liquid medium with 50 m m of MES containing 2% (w/v) Gal or 2% (w/v) Glc (as a negative control) for 2 h, followed by washing three times with sterilized Milli-Q water (EMD Millipore).

Plasmid Preparation:

Article Title: The ZIP Transporter Family Member OsZIP9 Contributes To Root Zinc Uptake in Rice under Zinc-Limited Conditions 1The ZIP Transporter Family Member OsZIP9 Contributes To Root Zinc Uptake in Rice under Zinc-Limited Conditions 1 [OPEN]
Article Snippet: .. In a time-course experiment, ZHY3 expressing OsZIP9 or empty vector was grown in the Sc(-Uracil) medium containing 0.67% (w/v) yeast nitrogen base without amino acids (Difco), 2% (w/v) Glc, 0.2% (w/v) appropriate amino acid, and 2% (w/v) agar at pH 6.0 for selection. .. The yeast cells were first incubated in Sc(-Uracil) liquid medium with 50 m m of MES containing 2% (w/v) Gal or 2% (w/v) Glc (as a negative control) for 2 h, followed by washing three times with sterilized Milli-Q water (EMD Millipore).

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    Difco amino acid auxotrophy experiments m9 minimal media
    Multi-spacer CRISPR arrays direct multiplex insertions in a single step. a, Schematic of multiplexed RNA-guided DNA integration events with pSPIN encoding a multi-spacer CRISPR array. b, qPCR-based quantification of integration efficiency with crRNA-NT (grey) and crRNA-4 (maroon), encoded in a single-, double-, or triple-spacer CRISPR array in the position indicated; white squares represent other functional crRNAs. Data are normalized to the single-spacer array efficiency. c, Tn-seq data for a triple-spacer CRISPR array, plotted as the percent of total genome-mapping reads. The target sites are denoted by colored triangles, and the insets show the distribution of integration events within a 42–58 bp window downstream of the target site. d, Schematic of experiment using thrC - and lysA -specific spacers for single-step generation of threonine-lysine auxotrophic E. coli . e, Recovery percentage of the indicated clonal genotypes (WT, single-knockout, or double-knockout) after transforming E. coli with pSPIN encoding a double-spacer CRISPR array containing both thrC - and lysA -specific spacers. Colonies were stamped onto M9-agar supplemented with either H 2 O, threonine, lysine, or threonine and lysine, and genotypes were determined based on survivability in each of these conditions. f, Growth curves for WT and double-knockout E. coli clones cultured at 37 °C in LB or <t>M9</t> minimal media with or without supplemented threonine (T) and lysine (L). g, Schematic of experimental approach to generate programmed genomic deletions. A double-spacer array directs multiplex insertion of two mini-Tn copies carrying LoxP sites; subsequent introduction of Cre recombinase leads to precise excision of the genomic fragment spanning the LoxP sites. h, Left, schematic showing genomic locus targeted for deletion. Right, the indicated double-spacer arrays were used to generate defined deletions, and PCR was performed with the indicated primer pairs to detect the presence/absence of genomic fragments flanking each target site, resolved by agarose gel electrophoresis. i, The programmed genomic deletions generated in h (2.4-, 10-, or 20-kb in length) were further verified by whole-genome, single-molecule real-time (SMRT) sequencing ( Methods ). The mini-transposon is indicated with a red arrow. Data in b and e are shown as mean ± s.d. for n = 3 biologically independent samples. Data in f are shown as mean ± s.d. for three technical replicates.
    Amino Acid Auxotrophy Experiments M9 Minimal Media, supplied by Difco, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/amino acid auxotrophy experiments m9 minimal media/product/Difco
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    85
    Difco synthetic drop out amino acid
    Multi-spacer CRISPR arrays direct multiplex insertions in a single step. a, Schematic of multiplexed RNA-guided DNA integration events with pSPIN encoding a multi-spacer CRISPR array. b, qPCR-based quantification of integration efficiency with crRNA-NT (grey) and crRNA-4 (maroon), encoded in a single-, double-, or triple-spacer CRISPR array in the position indicated; white squares represent other functional crRNAs. Data are normalized to the single-spacer array efficiency. c, Tn-seq data for a triple-spacer CRISPR array, plotted as the percent of total genome-mapping reads. The target sites are denoted by colored triangles, and the insets show the distribution of integration events within a 42–58 bp window downstream of the target site. d, Schematic of experiment using thrC - and lysA -specific spacers for single-step generation of threonine-lysine auxotrophic E. coli . e, Recovery percentage of the indicated clonal genotypes (WT, single-knockout, or double-knockout) after transforming E. coli with pSPIN encoding a double-spacer CRISPR array containing both thrC - and lysA -specific spacers. Colonies were stamped onto M9-agar supplemented with either H 2 O, threonine, lysine, or threonine and lysine, and genotypes were determined based on survivability in each of these conditions. f, Growth curves for WT and double-knockout E. coli clones cultured at 37 °C in LB or <t>M9</t> minimal media with or without supplemented threonine (T) and lysine (L). g, Schematic of experimental approach to generate programmed genomic deletions. A double-spacer array directs multiplex insertion of two mini-Tn copies carrying LoxP sites; subsequent introduction of Cre recombinase leads to precise excision of the genomic fragment spanning the LoxP sites. h, Left, schematic showing genomic locus targeted for deletion. Right, the indicated double-spacer arrays were used to generate defined deletions, and PCR was performed with the indicated primer pairs to detect the presence/absence of genomic fragments flanking each target site, resolved by agarose gel electrophoresis. i, The programmed genomic deletions generated in h (2.4-, 10-, or 20-kb in length) were further verified by whole-genome, single-molecule real-time (SMRT) sequencing ( Methods ). The mini-transposon is indicated with a red arrow. Data in b and e are shown as mean ± s.d. for n = 3 biologically independent samples. Data in f are shown as mean ± s.d. for three technical replicates.
    Synthetic Drop Out Amino Acid, supplied by Difco, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/synthetic drop out amino acid/product/Difco
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    80
    Difco amino acid free difco yeast nitrogen base
    Multi-spacer CRISPR arrays direct multiplex insertions in a single step. a, Schematic of multiplexed RNA-guided DNA integration events with pSPIN encoding a multi-spacer CRISPR array. b, qPCR-based quantification of integration efficiency with crRNA-NT (grey) and crRNA-4 (maroon), encoded in a single-, double-, or triple-spacer CRISPR array in the position indicated; white squares represent other functional crRNAs. Data are normalized to the single-spacer array efficiency. c, Tn-seq data for a triple-spacer CRISPR array, plotted as the percent of total genome-mapping reads. The target sites are denoted by colored triangles, and the insets show the distribution of integration events within a 42–58 bp window downstream of the target site. d, Schematic of experiment using thrC - and lysA -specific spacers for single-step generation of threonine-lysine auxotrophic E. coli . e, Recovery percentage of the indicated clonal genotypes (WT, single-knockout, or double-knockout) after transforming E. coli with pSPIN encoding a double-spacer CRISPR array containing both thrC - and lysA -specific spacers. Colonies were stamped onto M9-agar supplemented with either H 2 O, threonine, lysine, or threonine and lysine, and genotypes were determined based on survivability in each of these conditions. f, Growth curves for WT and double-knockout E. coli clones cultured at 37 °C in LB or <t>M9</t> minimal media with or without supplemented threonine (T) and lysine (L). g, Schematic of experimental approach to generate programmed genomic deletions. A double-spacer array directs multiplex insertion of two mini-Tn copies carrying LoxP sites; subsequent introduction of Cre recombinase leads to precise excision of the genomic fragment spanning the LoxP sites. h, Left, schematic showing genomic locus targeted for deletion. Right, the indicated double-spacer arrays were used to generate defined deletions, and PCR was performed with the indicated primer pairs to detect the presence/absence of genomic fragments flanking each target site, resolved by agarose gel electrophoresis. i, The programmed genomic deletions generated in h (2.4-, 10-, or 20-kb in length) were further verified by whole-genome, single-molecule real-time (SMRT) sequencing ( Methods ). The mini-transposon is indicated with a red arrow. Data in b and e are shown as mean ± s.d. for n = 3 biologically independent samples. Data in f are shown as mean ± s.d. for three technical replicates.
    Amino Acid Free Difco Yeast Nitrogen Base, supplied by Difco, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Difco 12 amino dodecanoic acid
    Multi-spacer CRISPR arrays direct multiplex insertions in a single step. a, Schematic of multiplexed RNA-guided DNA integration events with pSPIN encoding a multi-spacer CRISPR array. b, qPCR-based quantification of integration efficiency with crRNA-NT (grey) and crRNA-4 (maroon), encoded in a single-, double-, or triple-spacer CRISPR array in the position indicated; white squares represent other functional crRNAs. Data are normalized to the single-spacer array efficiency. c, Tn-seq data for a triple-spacer CRISPR array, plotted as the percent of total genome-mapping reads. The target sites are denoted by colored triangles, and the insets show the distribution of integration events within a 42–58 bp window downstream of the target site. d, Schematic of experiment using thrC - and lysA -specific spacers for single-step generation of threonine-lysine auxotrophic E. coli . e, Recovery percentage of the indicated clonal genotypes (WT, single-knockout, or double-knockout) after transforming E. coli with pSPIN encoding a double-spacer CRISPR array containing both thrC - and lysA -specific spacers. Colonies were stamped onto M9-agar supplemented with either H 2 O, threonine, lysine, or threonine and lysine, and genotypes were determined based on survivability in each of these conditions. f, Growth curves for WT and double-knockout E. coli clones cultured at 37 °C in LB or <t>M9</t> minimal media with or without supplemented threonine (T) and lysine (L). g, Schematic of experimental approach to generate programmed genomic deletions. A double-spacer array directs multiplex insertion of two mini-Tn copies carrying LoxP sites; subsequent introduction of Cre recombinase leads to precise excision of the genomic fragment spanning the LoxP sites. h, Left, schematic showing genomic locus targeted for deletion. Right, the indicated double-spacer arrays were used to generate defined deletions, and PCR was performed with the indicated primer pairs to detect the presence/absence of genomic fragments flanking each target site, resolved by agarose gel electrophoresis. i, The programmed genomic deletions generated in h (2.4-, 10-, or 20-kb in length) were further verified by whole-genome, single-molecule real-time (SMRT) sequencing ( Methods ). The mini-transposon is indicated with a red arrow. Data in b and e are shown as mean ± s.d. for n = 3 biologically independent samples. Data in f are shown as mean ± s.d. for three technical replicates.
    12 Amino Dodecanoic Acid, supplied by Difco, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Multi-spacer CRISPR arrays direct multiplex insertions in a single step. a, Schematic of multiplexed RNA-guided DNA integration events with pSPIN encoding a multi-spacer CRISPR array. b, qPCR-based quantification of integration efficiency with crRNA-NT (grey) and crRNA-4 (maroon), encoded in a single-, double-, or triple-spacer CRISPR array in the position indicated; white squares represent other functional crRNAs. Data are normalized to the single-spacer array efficiency. c, Tn-seq data for a triple-spacer CRISPR array, plotted as the percent of total genome-mapping reads. The target sites are denoted by colored triangles, and the insets show the distribution of integration events within a 42–58 bp window downstream of the target site. d, Schematic of experiment using thrC - and lysA -specific spacers for single-step generation of threonine-lysine auxotrophic E. coli . e, Recovery percentage of the indicated clonal genotypes (WT, single-knockout, or double-knockout) after transforming E. coli with pSPIN encoding a double-spacer CRISPR array containing both thrC - and lysA -specific spacers. Colonies were stamped onto M9-agar supplemented with either H 2 O, threonine, lysine, or threonine and lysine, and genotypes were determined based on survivability in each of these conditions. f, Growth curves for WT and double-knockout E. coli clones cultured at 37 °C in LB or M9 minimal media with or without supplemented threonine (T) and lysine (L). g, Schematic of experimental approach to generate programmed genomic deletions. A double-spacer array directs multiplex insertion of two mini-Tn copies carrying LoxP sites; subsequent introduction of Cre recombinase leads to precise excision of the genomic fragment spanning the LoxP sites. h, Left, schematic showing genomic locus targeted for deletion. Right, the indicated double-spacer arrays were used to generate defined deletions, and PCR was performed with the indicated primer pairs to detect the presence/absence of genomic fragments flanking each target site, resolved by agarose gel electrophoresis. i, The programmed genomic deletions generated in h (2.4-, 10-, or 20-kb in length) were further verified by whole-genome, single-molecule real-time (SMRT) sequencing ( Methods ). The mini-transposon is indicated with a red arrow. Data in b and e are shown as mean ± s.d. for n = 3 biologically independent samples. Data in f are shown as mean ± s.d. for three technical replicates.

    Journal: bioRxiv

    Article Title: CRISPR RNA-guided integrases for high-efficiency and multiplexed bacterial genome engineering

    doi: 10.1101/2020.07.17.209452

    Figure Lengend Snippet: Multi-spacer CRISPR arrays direct multiplex insertions in a single step. a, Schematic of multiplexed RNA-guided DNA integration events with pSPIN encoding a multi-spacer CRISPR array. b, qPCR-based quantification of integration efficiency with crRNA-NT (grey) and crRNA-4 (maroon), encoded in a single-, double-, or triple-spacer CRISPR array in the position indicated; white squares represent other functional crRNAs. Data are normalized to the single-spacer array efficiency. c, Tn-seq data for a triple-spacer CRISPR array, plotted as the percent of total genome-mapping reads. The target sites are denoted by colored triangles, and the insets show the distribution of integration events within a 42–58 bp window downstream of the target site. d, Schematic of experiment using thrC - and lysA -specific spacers for single-step generation of threonine-lysine auxotrophic E. coli . e, Recovery percentage of the indicated clonal genotypes (WT, single-knockout, or double-knockout) after transforming E. coli with pSPIN encoding a double-spacer CRISPR array containing both thrC - and lysA -specific spacers. Colonies were stamped onto M9-agar supplemented with either H 2 O, threonine, lysine, or threonine and lysine, and genotypes were determined based on survivability in each of these conditions. f, Growth curves for WT and double-knockout E. coli clones cultured at 37 °C in LB or M9 minimal media with or without supplemented threonine (T) and lysine (L). g, Schematic of experimental approach to generate programmed genomic deletions. A double-spacer array directs multiplex insertion of two mini-Tn copies carrying LoxP sites; subsequent introduction of Cre recombinase leads to precise excision of the genomic fragment spanning the LoxP sites. h, Left, schematic showing genomic locus targeted for deletion. Right, the indicated double-spacer arrays were used to generate defined deletions, and PCR was performed with the indicated primer pairs to detect the presence/absence of genomic fragments flanking each target site, resolved by agarose gel electrophoresis. i, The programmed genomic deletions generated in h (2.4-, 10-, or 20-kb in length) were further verified by whole-genome, single-molecule real-time (SMRT) sequencing ( Methods ). The mini-transposon is indicated with a red arrow. Data in b and e are shown as mean ± s.d. for n = 3 biologically independent samples. Data in f are shown as mean ± s.d. for three technical replicates.

    Article Snippet: Amino acid auxotrophy experiments M9 minimal media was prepared with the following components: 1X M9 salts (Difco), 0.4% glucose, 2 mM MgSO4 , and 0.1 mM CaCl2.

    Techniques: CRISPR, Multiplex Assay, Real-time Polymerase Chain Reaction, Functional Assay, Knock-Out, Double Knockout, Clone Assay, Cell Culture, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Generated, Sequencing