polymerase chain reaction pcr buffer ii  (Thermo Fisher)


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    Name:
    Platinum II Green PCR Buffer 5X
    Description:
    Invitrogen Platinum II Green PCR Buffer is an innovative buffer optimized for use with Platinum II Taq Hot Start DNA Polymerase Due to its unique composition the annealing temperature is 60°C for most primer pairs designed following the general design rules Isostabilizing molecules in the buffer increase primer template duplex stability during the annealing step and contribute to enhanced specificity without the need to optimize annealing temperature for each primer pair Platinum II Green PCR Buffer provides 1 5 mM MgCl2 in the final reaction and is supplemented with a density reagent and two tracking dyes for direct loading of PCR products on gels Find out more at www thermofisher com platinumiitaq ›
    Catalog Number:
    14966123
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    Category:
    Lab Reagents and Chemicals
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    Structured Review

    Thermo Fisher polymerase chain reaction pcr buffer ii
    Invitrogen Platinum II Green PCR Buffer is an innovative buffer optimized for use with Platinum II Taq Hot Start DNA Polymerase Due to its unique composition the annealing temperature is 60°C for most primer pairs designed following the general design rules Isostabilizing molecules in the buffer increase primer template duplex stability during the annealing step and contribute to enhanced specificity without the need to optimize annealing temperature for each primer pair Platinum II Green PCR Buffer provides 1 5 mM MgCl2 in the final reaction and is supplemented with a density reagent and two tracking dyes for direct loading of PCR products on gels Find out more at www thermofisher com platinumiitaq ›
    https://www.bioz.com/result/polymerase chain reaction pcr buffer ii/product/Thermo Fisher
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction pcr buffer ii - by Bioz Stars, 2020-07
    94/100 stars

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    Polymerase Chain Reaction:

    Article Title: Altered Subcellular Localization of Heat Shock Protein 90 Is Associated with Impaired Expression of the Aryl Hydrocarbon Receptor Pathway in Dogs
    Article Snippet: .. Standard amplification was performed using PCR-mix containing 1× Platinum® PCR Buffer, dNTPs 0.5 µM each, 2 mM MgCl2 , 0.5 mM of each forward and reverse primer, 1 Unit Platinum® Taq DNA polymerase (Invitrogen, Carlsbad, CA) and 50 ng of gDNA in a reaction volume of 25 µl. .. The thermal cycling protocol consisted of a 5-min denaturation step at 95°C, 35 cycles of 30 s at 95°C, 30 s at annealing temperature, 30 s at 72°C, and a final elongation step at 72°C for 10 min. For amplification of fragments exceeding 2000 bp, Phusion ™ Hot Start High-Fidelity DNA Polymerase (Thermo Scientific, Lafayette, US) was used.

    Article Title: Leaf endophytic fungus interacts with precipitation to alter belowground microbial communities in primary successional dunes
    Article Snippet: .. Briefly, PCRs were performed in triplicate 25 μL reactions containing 0.25 mM forward and reverse fusion primer, 0.25 mM dNTP (each), 1x Platinum PCR buffer (Lifetech, Carlsbad, CA), 1.5 mM MgCl2 , 1 U Platinum Taq Polymerase (Lifetech, Carlsbad, CA) and 2 μL (∼40 ng) of DNA template. .. Fusion primers were designed so that the forward primer consisted of the Roche adapter A, followed by a 10 base error-correcting barcode for multiplexing (Hamady et al. ), and using FLR3 (5΄-TTG AAA GGG AAA CGA TTG AAG T-3΄).

    Article Title: MicroRNA-98-5p regulates the proliferation and apoptosis of A549 cells by targeting MAP4K3
    Article Snippet: .. PCR was performed using the following thermocycling conditions: 94°C for 2 min, followed by 35 cycles of 94°C for 2 sec, 60°C for 60 sec and 72°C for 1 min with the following primers: MAP4K3 3′UTR forward, 5′-GGTACCAAAATAATTTAGTTACT-3′ and reverse, 5′-CTCGAGTGAGGTAGTAACT-3′ and Platinum™ II Green PCR buffer (Thermo Fisher Scientific, Inc.). .. The PCR products were amplified using cDNA from 293T cells and fused to the firefly luciferase gene of the pGL3-control plasmid (Promega Corporation) with the restriction enzyme sites of Kpn I and Xho I.

    Article Title: Optimisation of biomass, exopolysaccharide and intracellular polysaccharide production from the mycelium of an identified Ganoderma lucidum strain QRS 5120 using response surface methodology
    Article Snippet: .. Then, 0.5 pmol of ITS1 and ITS4 was added following by deoxynucleotide triphosphates (dNTPs, 200 μM each) (Promega no. U1511, Madison, OH, USA), 0.5 U DNA polymerase (Promega no. D1501, Madison, OH, USA), supplied PCR buffer (ThermoFisher no. 14966123, Platinum II Green PCR Buffer) and water. .. PCR was performed as follow: 1 cycle (98 °C for 2 min) for initial denaturation; 25 cycles (98 °C for 15 secs; 60 °C for 30 secs; 72 °C for 30 sec) for annealing and extension, and 1 cycle (72 °C for 10 min) for final extension of the amplified DNA (Eppendorf Mastercycler gradient, Hamburg, Germany) .

    Amplification:

    Article Title: Altered Subcellular Localization of Heat Shock Protein 90 Is Associated with Impaired Expression of the Aryl Hydrocarbon Receptor Pathway in Dogs
    Article Snippet: .. Standard amplification was performed using PCR-mix containing 1× Platinum® PCR Buffer, dNTPs 0.5 µM each, 2 mM MgCl2 , 0.5 mM of each forward and reverse primer, 1 Unit Platinum® Taq DNA polymerase (Invitrogen, Carlsbad, CA) and 50 ng of gDNA in a reaction volume of 25 µl. .. The thermal cycling protocol consisted of a 5-min denaturation step at 95°C, 35 cycles of 30 s at 95°C, 30 s at annealing temperature, 30 s at 72°C, and a final elongation step at 72°C for 10 min. For amplification of fragments exceeding 2000 bp, Phusion ™ Hot Start High-Fidelity DNA Polymerase (Thermo Scientific, Lafayette, US) was used.

    Size-exclusion Chromatography:

    Article Title: MicroRNA-98-5p regulates the proliferation and apoptosis of A549 cells by targeting MAP4K3
    Article Snippet: .. PCR was performed using the following thermocycling conditions: 94°C for 2 min, followed by 35 cycles of 94°C for 2 sec, 60°C for 60 sec and 72°C for 1 min with the following primers: MAP4K3 3′UTR forward, 5′-GGTACCAAAATAATTTAGTTACT-3′ and reverse, 5′-CTCGAGTGAGGTAGTAACT-3′ and Platinum™ II Green PCR buffer (Thermo Fisher Scientific, Inc.). .. The PCR products were amplified using cDNA from 293T cells and fused to the firefly luciferase gene of the pGL3-control plasmid (Promega Corporation) with the restriction enzyme sites of Kpn I and Xho I.

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    Thermo Fisher dna digestion mix
    Overview of RT-RamDA and single-cell RamDA-seq. a Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a RNA template. 2. Complementary <t>DNA</t> (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of RNase H minus reverse transcriptase (RTase). 3. Endonuclease <t>(DNase</t> I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the T4 gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I. b Relative yield of cDNA molecules using RT-qPCR ( n = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog , Pou5f1 , Zfp42 , and Sox2 ) and three housekeeping ( Gnb2l1 , Atp5a1 , and Tubb5 ) genes using a conventional method (−) as a standard. c Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section. d Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in b and d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile. e Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in d and e . Transcripts were annotated by GENCODE gene annotation (vM9)
    Dna Digestion Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna digestion mix/product/Thermo Fisher
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    Thermo Fisher dnase i reaction buffer
    Overview of RT-RamDA and single-cell RamDA-seq.  a  Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a RNA template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of RNase H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the T4 gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I.  b  Relative yield of cDNA molecules using RT-qPCR ( n  = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog ,  Pou5f1 ,  Zfp42 , and  Sox2 ) and three housekeeping ( Gnb2l1 ,  Atp5a1 , and  Tubb5 ) genes using a conventional method (−) as a standard.  c  Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section.  d  Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in  b  and  d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile.  e  Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in  d  and  e . Transcripts were annotated by GENCODE gene annotation (vM9)
    Dnase I Reaction Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dnase i reaction buffer/product/Thermo Fisher
    Average 99 stars, based on 25 article reviews
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    dnase i reaction buffer - by Bioz Stars, 2020-07
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    99
    Thermo Fisher dnase i solution
    Phosphoric acid is the most effective reagent for eliminating residual DNA from used Qiagen PCR purification kit columns. ( A ) A scheme showing the workflow for different protocols used in this study. ( B ) The standard curve that was plotted with the Ct values against –Log 10 DNA concentration (ng/μL). Ten microliters of LIF DNA template at a concentration of 100 ng/μL was consecutively diluted at 1/10 for 8 times, and the diluted samples (starting from 1 * 10 −2 ng/μL) were used for qPCR. Ct: Threshold cycles in quantitative PCR. ( C ) qPCR analysis to assess the amount of residual DNA eluted from the columns that were cleaned with different chemical reagents. LIF-contaminated columns were cleaned with indicated reagent based on protocol I. One microlitre of eluate was used to assess the presence of residual DNA by qPCR, with each sample assayed in triplicate. The obtained Ct values were converted into DNA concentration using the standard curve in B and were subjected to statistical analysis. The value of each column within the graph is presented in the bottom of the panel to clearly show the differences between the samples. ( D ) qPCR results for the amount of residual DNA from the columns that were cleaned with <t>DNase</t> I or acidic phenol. Contaminated columns were cleaned based on protocol II or III. qPCR assays were performed as in C. (E) qPCR showing DNA elimination for the used columns cleaned with different concentrations of phosphoric acid. Contaminated columns were cleaned with different concentrations of phosphoric acid as detailed in protocol I. qPCR assays performed as in C. Each column in C–E represents the mean ± SD of three independent experiments. * p
    Dnase I Solution, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Overview of RT-RamDA and single-cell RamDA-seq. a Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a RNA template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of RNase H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the T4 gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I. b Relative yield of cDNA molecules using RT-qPCR ( n = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog , Pou5f1 , Zfp42 , and Sox2 ) and three housekeeping ( Gnb2l1 , Atp5a1 , and Tubb5 ) genes using a conventional method (−) as a standard. c Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section. d Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in b and d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile. e Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in d and e . Transcripts were annotated by GENCODE gene annotation (vM9)

    Journal: Nature Communications

    Article Title: Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs

    doi: 10.1038/s41467-018-02866-0

    Figure Lengend Snippet: Overview of RT-RamDA and single-cell RamDA-seq. a Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a RNA template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of RNase H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the T4 gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I. b Relative yield of cDNA molecules using RT-qPCR ( n = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog , Pou5f1 , Zfp42 , and Sox2 ) and three housekeeping ( Gnb2l1 , Atp5a1 , and Tubb5 ) genes using a conventional method (−) as a standard. c Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section. d Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in b and d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile. e Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in d and e . Transcripts were annotated by GENCODE gene annotation (vM9)

    Article Snippet: To eliminate genomic DNA contamination, 1 μL of genomic DNA digestion mix (0.5× PrimeScript Buffer, 0.2 U of DNase I Amplification Grade, 1: 5 000 000 ERCC RNA Spike-In Mix I (Thermo Fisher) in RNase-free water) was added to 1 μL of the denatured sample.

    Techniques: Synthesized, Activity Assay, Amplification, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Expressing, Whisker Assay

    Overview of RT-RamDA and single-cell RamDA-seq.  a  Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a RNA template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of RNase H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the T4 gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I.  b  Relative yield of cDNA molecules using RT-qPCR ( n  = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog ,  Pou5f1 ,  Zfp42 , and  Sox2 ) and three housekeeping ( Gnb2l1 ,  Atp5a1 , and  Tubb5 ) genes using a conventional method (−) as a standard.  c  Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section.  d  Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in  b  and  d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile.  e  Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in  d  and  e . Transcripts were annotated by GENCODE gene annotation (vM9)

    Journal: Nature Communications

    Article Title: Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs

    doi: 10.1038/s41467-018-02866-0

    Figure Lengend Snippet: Overview of RT-RamDA and single-cell RamDA-seq. a Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a RNA template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of RNase H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the T4 gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I. b Relative yield of cDNA molecules using RT-qPCR ( n  = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog , Pou5f1 , Zfp42 , and Sox2 ) and three housekeeping ( Gnb2l1 , Atp5a1 , and Tubb5 ) genes using a conventional method (−) as a standard. c Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section. d Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in b and d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile. e Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in d and e . Transcripts were annotated by GENCODE gene annotation (vM9)

    Article Snippet: Next, 0.5 μL of genomic DNA digestion mix (0.1 U of DNase I Amplification Grade (Thermo Fisher) and 2× DNase I Reaction Buffer (Thermo Fisher) in RNase-free water) was added to 1 μL of the single-cell lysate in a 96-well PCR plate and incubated at 25 °C for 5 min. After genomic DNA digestion, we added 0.5 µL of denaturing mix (8 mM EDTA and 0.02% NP40 in RNase-free water) to the digested sample, followed by incubation at 70 °C for 5 min to inactivate DNase I and desaturate the RNAs.

    Techniques: Synthesized, Activity Assay, Amplification, Quantitative RT-PCR, Real-time Polymerase Chain Reaction, Expressing, Whisker Assay

    Phosphoric acid is the most effective reagent for eliminating residual DNA from used Qiagen PCR purification kit columns. ( A ) A scheme showing the workflow for different protocols used in this study. ( B ) The standard curve that was plotted with the Ct values against –Log 10 DNA concentration (ng/μL). Ten microliters of LIF DNA template at a concentration of 100 ng/μL was consecutively diluted at 1/10 for 8 times, and the diluted samples (starting from 1 * 10 −2 ng/μL) were used for qPCR. Ct: Threshold cycles in quantitative PCR. ( C ) qPCR analysis to assess the amount of residual DNA eluted from the columns that were cleaned with different chemical reagents. LIF-contaminated columns were cleaned with indicated reagent based on protocol I. One microlitre of eluate was used to assess the presence of residual DNA by qPCR, with each sample assayed in triplicate. The obtained Ct values were converted into DNA concentration using the standard curve in B and were subjected to statistical analysis. The value of each column within the graph is presented in the bottom of the panel to clearly show the differences between the samples. ( D ) qPCR results for the amount of residual DNA from the columns that were cleaned with DNase I or acidic phenol. Contaminated columns were cleaned based on protocol II or III. qPCR assays were performed as in C. (E) qPCR showing DNA elimination for the used columns cleaned with different concentrations of phosphoric acid. Contaminated columns were cleaned with different concentrations of phosphoric acid as detailed in protocol I. qPCR assays performed as in C. Each column in C–E represents the mean ± SD of three independent experiments. * p

    Journal: Scientific Reports

    Article Title: Rapid Regeneration and Reuse of Silica Columns from PCR Purification and Gel Extraction Kits

    doi: 10.1038/s41598-018-30316-w

    Figure Lengend Snippet: Phosphoric acid is the most effective reagent for eliminating residual DNA from used Qiagen PCR purification kit columns. ( A ) A scheme showing the workflow for different protocols used in this study. ( B ) The standard curve that was plotted with the Ct values against –Log 10 DNA concentration (ng/μL). Ten microliters of LIF DNA template at a concentration of 100 ng/μL was consecutively diluted at 1/10 for 8 times, and the diluted samples (starting from 1 * 10 −2 ng/μL) were used for qPCR. Ct: Threshold cycles in quantitative PCR. ( C ) qPCR analysis to assess the amount of residual DNA eluted from the columns that were cleaned with different chemical reagents. LIF-contaminated columns were cleaned with indicated reagent based on protocol I. One microlitre of eluate was used to assess the presence of residual DNA by qPCR, with each sample assayed in triplicate. The obtained Ct values were converted into DNA concentration using the standard curve in B and were subjected to statistical analysis. The value of each column within the graph is presented in the bottom of the panel to clearly show the differences between the samples. ( D ) qPCR results for the amount of residual DNA from the columns that were cleaned with DNase I or acidic phenol. Contaminated columns were cleaned based on protocol II or III. qPCR assays were performed as in C. (E) qPCR showing DNA elimination for the used columns cleaned with different concentrations of phosphoric acid. Contaminated columns were cleaned with different concentrations of phosphoric acid as detailed in protocol I. qPCR assays performed as in C. Each column in C–E represents the mean ± SD of three independent experiments. * p

    Article Snippet: Briefly, after the used columns from commercial kits were washed with ddH2 O, 600 μL of DNase I solution (Thermo Scientific) was added into individual columns, followed by one or two incubations at 37 °C for varied durations.

    Techniques: Polymerase Chain Reaction, Purification, Concentration Assay, Real-time Polymerase Chain Reaction