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    Thermo Fisher qubit 2 0 fluorometer measurements
    a  3-BrPA potentiates Prima-1 toxicity against A549 cells in 5 mM glucose. A549 cells (4X10 3 ) were seeded in tissue culture 96 well plates in complete medium containing 20 mM glucose and 10% fetal bovine serum, then washed 3X with PBS and treated as indicated in each case, in medium supplemented with physiological 5 mM glucose, 2 mM glutamine and 5% dialyzed serum for 48 h. Relative proliferation /toxicity was assayed fluorometrically in octuplicate by the Alamar Blue method by quantitating the conversion of resazurin to fluorescent resorufin [  8 ]. This revealed that 50 μM Prima-1 cooperated with 3-BrPA rather than with CHC to suppress A549 cell growth.  b  Prima-1 decreases SLC2A1-GLUT1 in A549 cells. Sparse cells were seeded in 5 cm tissue culture plates (5  ×  10 5 cells per plate) in complete Dulbecco’s medium containing 20 mM glucose supplemented with 10% serum for 18 h, then washed 3X with PBS and treated as indicated in each case, in medium supplemented with physiological 5 mM glucose, 2 mM glutamine and 5% dialyzed serum whenever indicated (+) for 24 h. After RNA extraction with TRIZOL and quantification in a Qubit® 2.0 Fluorometer, cDNAs were prepared for end-point PCR analysis as indicated under Methods.essentially similar results were obtained in cells treated with Prima-1 in 5 mM glucose (not shown). Cells treated in parallel with those used for RNA analysis were used for GLUT1 protein immune blot [  40 ].  c  Prima-1 activates p21CDKN1A gene expression in A549 cells in 5 mM glucose. qPCR was used to determine relative expression of the p21CDK1N1 gene in control and treated cells, after RNA extraction, cDNA preparation and qPCR, as indicated under Methods. *denotes significance between treated cells relative to controls
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    a  3-BrPA potentiates Prima-1 toxicity against A549 cells in 5 mM glucose. A549 cells (4X10 3 ) were seeded in tissue culture 96 well plates in complete medium containing 20 mM glucose and 10% fetal bovine serum, then washed 3X with PBS and treated as indicated in each case, in medium supplemented with physiological 5 mM glucose, 2 mM glutamine and 5% dialyzed serum for 48 h. Relative proliferation /toxicity was assayed fluorometrically in octuplicate by the Alamar Blue method by quantitating the conversion of resazurin to fluorescent resorufin [  8 ]. This revealed that 50 μM Prima-1 cooperated with 3-BrPA rather than with CHC to suppress A549 cell growth.  b  Prima-1 decreases SLC2A1-GLUT1 in A549 cells. Sparse cells were seeded in 5 cm tissue culture plates (5  ×  10 5 cells per plate) in complete Dulbecco’s medium containing 20 mM glucose supplemented with 10% serum for 18 h, then washed 3X with PBS and treated as indicated in each case, in medium supplemented with physiological 5 mM glucose, 2 mM glutamine and 5% dialyzed serum whenever indicated (+) for 24 h. After RNA extraction with TRIZOL and quantification in a Qubit® 2.0 Fluorometer, cDNAs were prepared for end-point PCR analysis as indicated under Methods.essentially similar results were obtained in cells treated with Prima-1 in 5 mM glucose (not shown). Cells treated in parallel with those used for RNA analysis were used for GLUT1 protein immune blot [  40 ].  c  Prima-1 activates p21CDKN1A gene expression in A549 cells in 5 mM glucose. qPCR was used to determine relative expression of the p21CDK1N1 gene in control and treated cells, after RNA extraction, cDNA preparation and qPCR, as indicated under Methods. *denotes significance between treated cells relative to controls

    Journal: BMC Cancer

    Article Title: Hypoxic resistance of KRAS mutant tumor cells to 3-Bromopyruvate is counteracted by Prima-1 and reversed by N-acetylcysteine

    doi: 10.1186/s12885-016-2930-9

    Figure Lengend Snippet: a 3-BrPA potentiates Prima-1 toxicity against A549 cells in 5 mM glucose. A549 cells (4X10 3 ) were seeded in tissue culture 96 well plates in complete medium containing 20 mM glucose and 10% fetal bovine serum, then washed 3X with PBS and treated as indicated in each case, in medium supplemented with physiological 5 mM glucose, 2 mM glutamine and 5% dialyzed serum for 48 h. Relative proliferation /toxicity was assayed fluorometrically in octuplicate by the Alamar Blue method by quantitating the conversion of resazurin to fluorescent resorufin [ 8 ]. This revealed that 50 μM Prima-1 cooperated with 3-BrPA rather than with CHC to suppress A549 cell growth. b Prima-1 decreases SLC2A1-GLUT1 in A549 cells. Sparse cells were seeded in 5 cm tissue culture plates (5  ×  10 5 cells per plate) in complete Dulbecco’s medium containing 20 mM glucose supplemented with 10% serum for 18 h, then washed 3X with PBS and treated as indicated in each case, in medium supplemented with physiological 5 mM glucose, 2 mM glutamine and 5% dialyzed serum whenever indicated (+) for 24 h. After RNA extraction with TRIZOL and quantification in a Qubit® 2.0 Fluorometer, cDNAs were prepared for end-point PCR analysis as indicated under Methods.essentially similar results were obtained in cells treated with Prima-1 in 5 mM glucose (not shown). Cells treated in parallel with those used for RNA analysis were used for GLUT1 protein immune blot [ 40 ]. c Prima-1 activates p21CDKN1A gene expression in A549 cells in 5 mM glucose. qPCR was used to determine relative expression of the p21CDK1N1 gene in control and treated cells, after RNA extraction, cDNA preparation and qPCR, as indicated under Methods. *denotes significance between treated cells relative to controls

    Article Snippet: Cells were seeded in 5 cm-well plates (3 × 105 cells per plate) in complete Dulbecco’s medium containing 20 mM glucose supplemented with 10% serum for 24 h. Cells were washed 3X with PBS and treated as indicated in medium supplemented with physiological 5 mM glucose and 5% dialyzed serum for 24 h. RNA extraction was performed using TRIZOL® (Life Technologies, Cat # 15596–026) and quantification was determined using a Qubit® 2.0 Fluorometer (Life Technologies, Cat #Q32866) with a Qubit™ RNA Assay Kit (Life Technologies, Cat # Q32852).

    Techniques: RNA Extraction, Polymerase Chain Reaction, Expressing, Real-time Polymerase Chain Reaction

    Random hexamer primer detection by a dsDNA-specific fluorometric assay. Random hexamers (250, 500, 1000, 2000, 4000 ng) were added to 200 μL of the PureLyse elution buffer, homogenized, then quantified on a Qubit 2.0 fluorometer (standard error shown,  n  = 3). Random hexamer loads were not detectable until 20 ng/μL (4000 ng in 200 μL), likely due to increased hybridization. These results suggest that using random hexamer primers (less than 20 ng/μL) as competitive binders will not bias DNA yield results with this dsDNA-specific fluorometric assay. *At or below 0.01 ng limit of detection.

    Journal: Astrobiology

    Article Title: Nucleic Acid Extraction from Synthetic Mars Analog Soils for in situ Life Detection

    doi: 10.1089/ast.2016.1535

    Figure Lengend Snippet: Random hexamer primer detection by a dsDNA-specific fluorometric assay. Random hexamers (250, 500, 1000, 2000, 4000 ng) were added to 200 μL of the PureLyse elution buffer, homogenized, then quantified on a Qubit 2.0 fluorometer (standard error shown, n  = 3). Random hexamer loads were not detectable until 20 ng/μL (4000 ng in 200 μL), likely due to increased hybridization. These results suggest that using random hexamer primers (less than 20 ng/μL) as competitive binders will not bias DNA yield results with this dsDNA-specific fluorometric assay. *At or below 0.01 ng limit of detection.

    Article Snippet: DNAout was quantified using a double-stranded, DNA-specific fluorometric assay (Invitrogen, Qubit dsDNA HS Assay Kit, Q32854) and a Qubit 2.0 fluorometer (Invitrogen, Q32866, limit of detection of 0.1 ng/mL, dsDNA).

    Techniques: Random Hexamer Labeling, Hybridization

    PCR amplification and DNA concentration in decoction with sliced or pulverized sample at different times of boiling. (A) P. ginseng  sample was sliced (lanes 1–3, 7–9) or pulverized (lanes 4–6, 10–12) and boiled for 30 min (lanes 1, 4, 7, 10), 60 min (lanes 2, 5, 8, 11) and 120 min (lanes 3, 6, 9, 12). Crude DNA (lanes 1–6) and extracted DNA with concentration adjusted to that in the original decoction (lanes 7–12) were amplified by primer pair 9. Lane 13 is the positive control with DNA from  P. ginseng  and lane 14 is the negative control without DNA. Lane M represents the DNA size ladder.  (B)  The concentration of extracted  P. ginseng  DNA was adjusted to make it similar to that in the original decoction and measured by Qubit 2.0 Fluorometer. Water was used as blank and the data represent mean ± standard deviation (n = 3). Difference between boiling time was analyzed by one-way analysis of variance. Difference between sliced and pulverized samples at the same boiling time was analyzed by two sample  t  test (* p

    Journal: Chinese Medicine

    Article Title: Identification of constituent herbs in ginseng decoctions by DNA markers

    doi: 10.1186/s13020-015-0029-x

    Figure Lengend Snippet: PCR amplification and DNA concentration in decoction with sliced or pulverized sample at different times of boiling. (A) P. ginseng sample was sliced (lanes 1–3, 7–9) or pulverized (lanes 4–6, 10–12) and boiled for 30 min (lanes 1, 4, 7, 10), 60 min (lanes 2, 5, 8, 11) and 120 min (lanes 3, 6, 9, 12). Crude DNA (lanes 1–6) and extracted DNA with concentration adjusted to that in the original decoction (lanes 7–12) were amplified by primer pair 9. Lane 13 is the positive control with DNA from P. ginseng and lane 14 is the negative control without DNA. Lane M represents the DNA size ladder. (B) The concentration of extracted P. ginseng DNA was adjusted to make it similar to that in the original decoction and measured by Qubit 2.0 Fluorometer. Water was used as blank and the data represent mean ± standard deviation (n = 3). Difference between boiling time was analyzed by one-way analysis of variance. Difference between sliced and pulverized samples at the same boiling time was analyzed by two sample t test (* p

    Article Snippet: DNA concentration was measured using a Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA, USA) and presented as the mean ± standard deviation.

    Techniques: Polymerase Chain Reaction, Amplification, Concentration Assay, Positive Control, Negative Control, Standard Deviation

    Comparison of five automated DNA extraction systems. Illustration of relative DNA concentrations of samples 1–10 measured by the NanoDrop 2000c spectrophotometer (A) and Qubit 2.0 fluorometer (B). The concentrations of the Maxwell 16 extracts were set to 100% for each sample.

    Journal: PLoS ONE

    Article Title: Comparison of Pre-Analytical FFPE Sample Preparation Methods and Their Impact on Massively Parallel Sequencing in Routine Diagnostics

    doi: 10.1371/journal.pone.0104566

    Figure Lengend Snippet: Comparison of five automated DNA extraction systems. Illustration of relative DNA concentrations of samples 1–10 measured by the NanoDrop 2000c spectrophotometer (A) and Qubit 2.0 fluorometer (B). The concentrations of the Maxwell 16 extracts were set to 100% for each sample.

    Article Snippet: Additionally, each DNA sample was quantified in duplicates with the Qubit 2.0 fluorometer (Life Technologies, Darmstadt, Germany).

    Techniques: DNA Extraction, Spectrophotometry

    Cumulative DNA yields in successive extractions of clay (C1–C3) and sandy soils (S1–S3) using PS and FS DNA extraction kits. Average and error bars (SD) of all three biological replicates are presented for each DNA extraction. (A) Clay soils extracted with PS; (B) sandy soils extracted with PS; (C) clay soils extracted with FS; (D) sandy soils extracted with FS. DNA quantification was performed using a Qubit 2.0 fluorometer.

    Journal: PeerJ

    Article Title: Successive DNA extractions improve characterization of soil microbial communities

    doi: 10.7717/peerj.2915

    Figure Lengend Snippet: Cumulative DNA yields in successive extractions of clay (C1–C3) and sandy soils (S1–S3) using PS and FS DNA extraction kits. Average and error bars (SD) of all three biological replicates are presented for each DNA extraction. (A) Clay soils extracted with PS; (B) sandy soils extracted with PS; (C) clay soils extracted with FS; (D) sandy soils extracted with FS. DNA quantification was performed using a Qubit 2.0 fluorometer.

    Article Snippet: Total DNA quantity and quality were measured using a Qubit 2.0 fluorometer (Life Technologies, Carlsbad, CA, USA) and a NanoDrop 1000 spectrophotometer (Thermo Scientific, Wilmington, DE, USA), as well as visualized on 1% (w/v) agarose gel under UV light after staining with ethidium bromide.

    Techniques: DNA Extraction