pcr purification kit Search Results


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  • 99
    New England Biolabs polymerase chain reaction pcr clean kit
    Polymerase Chain Reaction Pcr Clean Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    tiangen biotech co polymerase chain reaction pcr purification kit
    Polymerase Chain Reaction Pcr Purification Kit, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Roche polymerase chain reaction pcr template purification kit
    Polymerase Chain Reaction Pcr Template Purification Kit, supplied by Roche, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen polymerase chain reaction pcr purification kit
    Polymerase Chain Reaction Pcr Purification Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 116 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Promega polymerase chain reaction pcr purification kit
    Polymerase Chain Reaction Pcr Purification Kit, supplied by Promega, used in various techniques. Bioz Stars score: 98/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen qiaquick polymerase chain reaction pcr purification kit
    Evaluation of TPW for different silica-column NA extraction kit protocols on pure water samples using ( a – c ) qPCR and ( d – f ) LAMP. All reactions were spiked with 5 × 10 4 copies λ phage DNA and primers. By manufacturer protocol, the ( a , d ) Zymo Quick-DNA/RNA Viral Kit and ( b , e ) Zymo ZR Viral DNA/RNA Kit do not include the dry spin (+dry spin) whereas the ( c , f ) Qiagen <t>QIAquick</t> <t>PCR</t> Purification Kit does. The left of each graph shows high dilution and the right shows low dilution. Each bar represents the result from a single qPCR or LAMP measurement. We ran 27 silica-column extractions (3 silica columns × 3 conditions × 3 extraction protocols) and the kit extract was shared between high and low dilutions of both qPCR and LAMP. Dashed lines show the C q or TTP for a reaction without inhibitors (“No Extract”). Samples marked N.D. were not detected within either 40 cycles or 40 min. ( a – f ) We asked whether the manufacturer protocol replicates (“No Dry Spin for Zymo kits, “+dry spin” for Qiagen kit) fell within the 95% CI of the corresponding +1-undecanol condition for the low kit extract dilution case. The number of replicates that lie outside the 95% CI are indicated by the number of + (above) and - (below).
    Qiaquick Polymerase Chain Reaction Pcr Purification Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 20135 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bioteke Corporation polymerase chain reaction pcr purification kit
    Evaluation of TPW for different silica-column NA extraction kit protocols on pure water samples using ( a – c ) qPCR and ( d – f ) LAMP. All reactions were spiked with 5 × 10 4 copies λ phage DNA and primers. By manufacturer protocol, the ( a , d ) Zymo Quick-DNA/RNA Viral Kit and ( b , e ) Zymo ZR Viral DNA/RNA Kit do not include the dry spin (+dry spin) whereas the ( c , f ) Qiagen <t>QIAquick</t> <t>PCR</t> Purification Kit does. The left of each graph shows high dilution and the right shows low dilution. Each bar represents the result from a single qPCR or LAMP measurement. We ran 27 silica-column extractions (3 silica columns × 3 conditions × 3 extraction protocols) and the kit extract was shared between high and low dilutions of both qPCR and LAMP. Dashed lines show the C q or TTP for a reaction without inhibitors (“No Extract”). Samples marked N.D. were not detected within either 40 cycles or 40 min. ( a – f ) We asked whether the manufacturer protocol replicates (“No Dry Spin for Zymo kits, “+dry spin” for Qiagen kit) fell within the 95% CI of the corresponding +1-undecanol condition for the low kit extract dilution case. The number of replicates that lie outside the 95% CI are indicated by the number of + (above) and - (below).
    Polymerase Chain Reaction Pcr Purification Kit, supplied by Bioteke Corporation, used in various techniques. Bioz Stars score: 92/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    iNtRON Biotechnology polymerase chain reaction pcr quick spin purification kit
    Evaluation of TPW for different silica-column NA extraction kit protocols on pure water samples using ( a – c ) qPCR and ( d – f ) LAMP. All reactions were spiked with 5 × 10 4 copies λ phage DNA and primers. By manufacturer protocol, the ( a , d ) Zymo Quick-DNA/RNA Viral Kit and ( b , e ) Zymo ZR Viral DNA/RNA Kit do not include the dry spin (+dry spin) whereas the ( c , f ) Qiagen <t>QIAquick</t> <t>PCR</t> Purification Kit does. The left of each graph shows high dilution and the right shows low dilution. Each bar represents the result from a single qPCR or LAMP measurement. We ran 27 silica-column extractions (3 silica columns × 3 conditions × 3 extraction protocols) and the kit extract was shared between high and low dilutions of both qPCR and LAMP. Dashed lines show the C q or TTP for a reaction without inhibitors (“No Extract”). Samples marked N.D. were not detected within either 40 cycles or 40 min. ( a – f ) We asked whether the manufacturer protocol replicates (“No Dry Spin for Zymo kits, “+dry spin” for Qiagen kit) fell within the 95% CI of the corresponding +1-undecanol condition for the low kit extract dilution case. The number of replicates that lie outside the 95% CI are indicated by the number of + (above) and - (below).
    Polymerase Chain Reaction Pcr Quick Spin Purification Kit, supplied by iNtRON Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polymerase chain reaction pcr quick spin purification kit/product/iNtRON Biotechnology
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    90
    Biomiga gel polymerase chain reaction pcr extraction kit
    Evaluation of TPW for different silica-column NA extraction kit protocols on pure water samples using ( a – c ) qPCR and ( d – f ) LAMP. All reactions were spiked with 5 × 10 4 copies λ phage DNA and primers. By manufacturer protocol, the ( a , d ) Zymo Quick-DNA/RNA Viral Kit and ( b , e ) Zymo ZR Viral DNA/RNA Kit do not include the dry spin (+dry spin) whereas the ( c , f ) Qiagen <t>QIAquick</t> <t>PCR</t> Purification Kit does. The left of each graph shows high dilution and the right shows low dilution. Each bar represents the result from a single qPCR or LAMP measurement. We ran 27 silica-column extractions (3 silica columns × 3 conditions × 3 extraction protocols) and the kit extract was shared between high and low dilutions of both qPCR and LAMP. Dashed lines show the C q or TTP for a reaction without inhibitors (“No Extract”). Samples marked N.D. were not detected within either 40 cycles or 40 min. ( a – f ) We asked whether the manufacturer protocol replicates (“No Dry Spin for Zymo kits, “+dry spin” for Qiagen kit) fell within the 95% CI of the corresponding +1-undecanol condition for the low kit extract dilution case. The number of replicates that lie outside the 95% CI are indicated by the number of + (above) and - (below).
    Gel Polymerase Chain Reaction Pcr Extraction Kit, supplied by Biomiga, used in various techniques. Bioz Stars score: 90/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Real Biotech Corporation hiyield polymerase chain reaction dna purification kit
    Evaluation of TPW for different silica-column NA extraction kit protocols on pure water samples using ( a – c ) qPCR and ( d – f ) LAMP. All reactions were spiked with 5 × 10 4 copies λ phage DNA and primers. By manufacturer protocol, the ( a , d ) Zymo Quick-DNA/RNA Viral Kit and ( b , e ) Zymo ZR Viral DNA/RNA Kit do not include the dry spin (+dry spin) whereas the ( c , f ) Qiagen <t>QIAquick</t> <t>PCR</t> Purification Kit does. The left of each graph shows high dilution and the right shows low dilution. Each bar represents the result from a single qPCR or LAMP measurement. We ran 27 silica-column extractions (3 silica columns × 3 conditions × 3 extraction protocols) and the kit extract was shared between high and low dilutions of both qPCR and LAMP. Dashed lines show the C q or TTP for a reaction without inhibitors (“No Extract”). Samples marked N.D. were not detected within either 40 cycles or 40 min. ( a – f ) We asked whether the manufacturer protocol replicates (“No Dry Spin for Zymo kits, “+dry spin” for Qiagen kit) fell within the 95% CI of the corresponding +1-undecanol condition for the low kit extract dilution case. The number of replicates that lie outside the 95% CI are indicated by the number of + (above) and - (below).
    Hiyield Polymerase Chain Reaction Dna Purification Kit, supplied by Real Biotech Corporation, used in various techniques. Bioz Stars score: 86/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche polymerase chain reaction pcr purification kits
    Quantitative analysis of vascular endothelial growth factor 2 ( VEGFR2 ) gene expression levels downregulated in AGS cells 72 hours after treatment with anti- VEGFR2 small interfering <t>RNA</t> ( siRNA ). Each real-time polymerase chain reaction <t>(PCR)</t> analysis was carried out at least in triplicate. Data are the fold change in relative expression compared with glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) based on the comparative Ct (2 -ΔΔCt ) method. Values are shown as mean ± SD.
    Polymerase Chain Reaction Pcr Purification Kits, supplied by Roche, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Gentra Systems polymerase chain reaction a genomic dna purification kit
    Quantitative analysis of vascular endothelial growth factor 2 ( VEGFR2 ) gene expression levels downregulated in AGS cells 72 hours after treatment with anti- VEGFR2 small interfering <t>RNA</t> ( siRNA ). Each real-time polymerase chain reaction <t>(PCR)</t> analysis was carried out at least in triplicate. Data are the fold change in relative expression compared with glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) based on the comparative Ct (2 -ΔΔCt ) method. Values are shown as mean ± SD.
    Polymerase Chain Reaction A Genomic Dna Purification Kit, supplied by Gentra Systems, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa polymerase chain reaction pcr purification kit
    Quantitative analysis of vascular endothelial growth factor 2 ( VEGFR2 ) gene expression levels downregulated in AGS cells 72 hours after treatment with anti- VEGFR2 small interfering <t>RNA</t> ( siRNA ). Each real-time polymerase chain reaction <t>(PCR)</t> analysis was carried out at least in triplicate. Data are the fold change in relative expression compared with glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) based on the comparative Ct (2 -ΔΔCt ) method. Values are shown as mean ± SD.
    Polymerase Chain Reaction Pcr Purification Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MACHEREY NAGEL polymerase chain reaction pcr clean up kit
    Quantitative analysis of vascular endothelial growth factor 2 ( VEGFR2 ) gene expression levels downregulated in AGS cells 72 hours after treatment with anti- VEGFR2 small interfering <t>RNA</t> ( siRNA ). Each real-time polymerase chain reaction <t>(PCR)</t> analysis was carried out at least in triplicate. Data are the fold change in relative expression compared with glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) based on the comparative Ct (2 -ΔΔCt ) method. Values are shown as mean ± SD.
    Polymerase Chain Reaction Pcr Clean Up Kit, supplied by MACHEREY NAGEL, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Qiagen minelute pcr purification kit
    DNA purification reagents vary in their ability to recover low amounts of DNA from de-crosslinked chromatin. a Recovered DNA amount by different DNA purification reagents from de-crosslinked chromatin. De-crosslinked chromatin estimated to include 1 ng range DNA in ChIP elution buffer was purified following the manufacturer’s instructions. The data were generated from triplicate DNA samples derived from three independent preparations. Zy, ChIP DNA Clean Concentrator™ (Zymo Research); Pr, Wizard® SV Gel and <t>PCR</t> Clean-Up System (Promega); Th, GeneJET PCR Purification Kit (Thermo Fisher Scientific); In, PureLink® PCR Purification Kit (Invitrogen); Ne, Monarch® PCR DNA Cleanup Kit (New England Biolabs); Am, Chromatin IP DNA Purification Kit (Active Motif); Qp, QIAquick PCR Purification Kit (Qiagen); Qm, <t>MinElute</t> PCR Purification Kit (Qiagen); Ba, Agencourt AMPure XP kit (Beckman, chromatin to beads ratio from 1:1.25 to 1:2); Br, RNAClean™ XP kit (Beckman, chromatin to beads ratio from 1:1.25 to 1:2); PC, phenol/chloroform extraction. b Interference of PCR amplification by purified eluent of purification reagents. 9 μL eluent was mixed with 1 μL 166 bp of Drosophila probe DNA (0.0001 ng), and the resulting mixture was used as the template in 20 μl of real-time PCR reaction. The Ct value for Drosophila probe DNA from TE buffer was set as 100%. The experiment was repeated 3 times using de-crosslinked chromatin estimated to include 1 ng of DNA. c Size profiles of DNA purified by different reagents. The DNAs purified from de-crosslinked chromatin estimated to include 50 ng range DNA was analyzed by AATI Fragment Analyzer. DNA size (bp) is shown
    Minelute Pcr Purification Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 21408 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene transcription polymerase chain reaction rt pcr kit
    Detection by reverse transcription–polymerase chain reaction <t>(RT–PCR)</t> of MUC1 transcript in activated T cells. (a) Human T cells were activated in vitro by anti-CD3 antibody. Total RNA was extracted and RT–PCR performed on the indicated days (D0, day 0; D1, day 1; D2, day 2; D3, day 3; D4, day 4). MUC1 transcript (446 bp) was identified after at least 24 hr. Human β-actin was included as a positive control, and a molecular weight marker is present in lane 1. (b) Semiquantitative RT–PCR for MUC1 in activated T cells and autologous breast cancer. Total RNA was extracted from autologous breast cancer cells and from purified human T cells following 4 days of in vitro activation using anti-CD3 antibody. <t>cDNA</t> was synthesized and then used as the template for RT–PCR at the dilutions shown. Human β-actin was included as a positive control.
    Transcription Polymerase Chain Reaction Rt Pcr Kit, supplied by Stratagene, used in various techniques. Bioz Stars score: 89/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    TaKaRa reverse transcription polymerase chain reaction rt pcr kit
    Detection by reverse transcription–polymerase chain reaction <t>(RT–PCR)</t> of MUC1 transcript in activated T cells. (a) Human T cells were activated in vitro by anti-CD3 antibody. Total RNA was extracted and RT–PCR performed on the indicated days (D0, day 0; D1, day 1; D2, day 2; D3, day 3; D4, day 4). MUC1 transcript (446 bp) was identified after at least 24 hr. Human β-actin was included as a positive control, and a molecular weight marker is present in lane 1. (b) Semiquantitative RT–PCR for MUC1 in activated T cells and autologous breast cancer. Total RNA was extracted from autologous breast cancer cells and from purified human T cells following 4 days of in vitro activation using anti-CD3 antibody. <t>cDNA</t> was synthesized and then used as the template for RT–PCR at the dilutions shown. Human β-actin was included as a positive control.
    Reverse Transcription Polymerase Chain Reaction Rt Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 136 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Axygen axyprep mag polymerase chain reaction pcr clean up kit
    Detection by reverse transcription–polymerase chain reaction <t>(RT–PCR)</t> of MUC1 transcript in activated T cells. (a) Human T cells were activated in vitro by anti-CD3 antibody. Total RNA was extracted and RT–PCR performed on the indicated days (D0, day 0; D1, day 1; D2, day 2; D3, day 3; D4, day 4). MUC1 transcript (446 bp) was identified after at least 24 hr. Human β-actin was included as a positive control, and a molecular weight marker is present in lane 1. (b) Semiquantitative RT–PCR for MUC1 in activated T cells and autologous breast cancer. Total RNA was extracted from autologous breast cancer cells and from purified human T cells following 4 days of in vitro activation using anti-CD3 antibody. <t>cDNA</t> was synthesized and then used as the template for RT–PCR at the dilutions shown. Human β-actin was included as a positive control.
    Axyprep Mag Polymerase Chain Reaction Pcr Clean Up Kit, supplied by Axygen, used in various techniques. Bioz Stars score: 93/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Evaluation of TPW for different silica-column NA extraction kit protocols on pure water samples using ( a – c ) qPCR and ( d – f ) LAMP. All reactions were spiked with 5 × 10 4 copies λ phage DNA and primers. By manufacturer protocol, the ( a , d ) Zymo Quick-DNA/RNA Viral Kit and ( b , e ) Zymo ZR Viral DNA/RNA Kit do not include the dry spin (+dry spin) whereas the ( c , f ) Qiagen QIAquick PCR Purification Kit does. The left of each graph shows high dilution and the right shows low dilution. Each bar represents the result from a single qPCR or LAMP measurement. We ran 27 silica-column extractions (3 silica columns × 3 conditions × 3 extraction protocols) and the kit extract was shared between high and low dilutions of both qPCR and LAMP. Dashed lines show the C q or TTP for a reaction without inhibitors (“No Extract”). Samples marked N.D. were not detected within either 40 cycles or 40 min. ( a – f ) We asked whether the manufacturer protocol replicates (“No Dry Spin for Zymo kits, “+dry spin” for Qiagen kit) fell within the 95% CI of the corresponding +1-undecanol condition for the low kit extract dilution case. The number of replicates that lie outside the 95% CI are indicated by the number of + (above) and - (below).

    Journal: Scientific Reports

    Article Title: Two-phase wash to solve the ubiquitous contaminant-carryover problem in commercial nucleic-acid extraction kits

    doi: 10.1038/s41598-020-58586-3

    Figure Lengend Snippet: Evaluation of TPW for different silica-column NA extraction kit protocols on pure water samples using ( a – c ) qPCR and ( d – f ) LAMP. All reactions were spiked with 5 × 10 4 copies λ phage DNA and primers. By manufacturer protocol, the ( a , d ) Zymo Quick-DNA/RNA Viral Kit and ( b , e ) Zymo ZR Viral DNA/RNA Kit do not include the dry spin (+dry spin) whereas the ( c , f ) Qiagen QIAquick PCR Purification Kit does. The left of each graph shows high dilution and the right shows low dilution. Each bar represents the result from a single qPCR or LAMP measurement. We ran 27 silica-column extractions (3 silica columns × 3 conditions × 3 extraction protocols) and the kit extract was shared between high and low dilutions of both qPCR and LAMP. Dashed lines show the C q or TTP for a reaction without inhibitors (“No Extract”). Samples marked N.D. were not detected within either 40 cycles or 40 min. ( a – f ) We asked whether the manufacturer protocol replicates (“No Dry Spin for Zymo kits, “+dry spin” for Qiagen kit) fell within the 95% CI of the corresponding +1-undecanol condition for the low kit extract dilution case. The number of replicates that lie outside the 95% CI are indicated by the number of + (above) and - (below).

    Article Snippet: Kit extractions We tested three different silica-column kits: Zymo ZR Viral DNA/RNA Kit (outdated protocol, D7021), Zymo Quick-DNA/RNA Kit (updated protocol, D7021), and the QIAquick PCR Purification Kit (28104, Qiagen).

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

    Quantitative analysis of vascular endothelial growth factor 2 ( VEGFR2 ) gene expression levels downregulated in AGS cells 72 hours after treatment with anti- VEGFR2 small interfering RNA ( siRNA ). Each real-time polymerase chain reaction (PCR) analysis was carried out at least in triplicate. Data are the fold change in relative expression compared with glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) based on the comparative Ct (2 -ΔΔCt ) method. Values are shown as mean ± SD.

    Journal: Cell Journal (Yakhteh)

    Article Title: Inhibition of AGS Cancer Cell Proliferation following siRNA-Mediated Downregulation of VEGFR2

    doi:

    Figure Lengend Snippet: Quantitative analysis of vascular endothelial growth factor 2 ( VEGFR2 ) gene expression levels downregulated in AGS cells 72 hours after treatment with anti- VEGFR2 small interfering RNA ( siRNA ). Each real-time polymerase chain reaction (PCR) analysis was carried out at least in triplicate. Data are the fold change in relative expression compared with glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) based on the comparative Ct (2 -ΔΔCt ) method. Values are shown as mean ± SD.

    Article Snippet: RNA extraction, cDNA synthesis and polymerase chain reaction (PCR) purification kits were obtained from Roche, Germany.

    Techniques: Expressing, Small Interfering RNA, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction

    DNA purification reagents vary in their ability to recover low amounts of DNA from de-crosslinked chromatin. a Recovered DNA amount by different DNA purification reagents from de-crosslinked chromatin. De-crosslinked chromatin estimated to include 1 ng range DNA in ChIP elution buffer was purified following the manufacturer’s instructions. The data were generated from triplicate DNA samples derived from three independent preparations. Zy, ChIP DNA Clean Concentrator™ (Zymo Research); Pr, Wizard® SV Gel and PCR Clean-Up System (Promega); Th, GeneJET PCR Purification Kit (Thermo Fisher Scientific); In, PureLink® PCR Purification Kit (Invitrogen); Ne, Monarch® PCR DNA Cleanup Kit (New England Biolabs); Am, Chromatin IP DNA Purification Kit (Active Motif); Qp, QIAquick PCR Purification Kit (Qiagen); Qm, MinElute PCR Purification Kit (Qiagen); Ba, Agencourt AMPure XP kit (Beckman, chromatin to beads ratio from 1:1.25 to 1:2); Br, RNAClean™ XP kit (Beckman, chromatin to beads ratio from 1:1.25 to 1:2); PC, phenol/chloroform extraction. b Interference of PCR amplification by purified eluent of purification reagents. 9 μL eluent was mixed with 1 μL 166 bp of Drosophila probe DNA (0.0001 ng), and the resulting mixture was used as the template in 20 μl of real-time PCR reaction. The Ct value for Drosophila probe DNA from TE buffer was set as 100%. The experiment was repeated 3 times using de-crosslinked chromatin estimated to include 1 ng of DNA. c Size profiles of DNA purified by different reagents. The DNAs purified from de-crosslinked chromatin estimated to include 50 ng range DNA was analyzed by AATI Fragment Analyzer. DNA size (bp) is shown

    Journal: BMC Genomics

    Article Title: Purification of nanogram-range immunoprecipitated DNA in ChIP-seq application

    doi: 10.1186/s12864-017-4371-5

    Figure Lengend Snippet: DNA purification reagents vary in their ability to recover low amounts of DNA from de-crosslinked chromatin. a Recovered DNA amount by different DNA purification reagents from de-crosslinked chromatin. De-crosslinked chromatin estimated to include 1 ng range DNA in ChIP elution buffer was purified following the manufacturer’s instructions. The data were generated from triplicate DNA samples derived from three independent preparations. Zy, ChIP DNA Clean Concentrator™ (Zymo Research); Pr, Wizard® SV Gel and PCR Clean-Up System (Promega); Th, GeneJET PCR Purification Kit (Thermo Fisher Scientific); In, PureLink® PCR Purification Kit (Invitrogen); Ne, Monarch® PCR DNA Cleanup Kit (New England Biolabs); Am, Chromatin IP DNA Purification Kit (Active Motif); Qp, QIAquick PCR Purification Kit (Qiagen); Qm, MinElute PCR Purification Kit (Qiagen); Ba, Agencourt AMPure XP kit (Beckman, chromatin to beads ratio from 1:1.25 to 1:2); Br, RNAClean™ XP kit (Beckman, chromatin to beads ratio from 1:1.25 to 1:2); PC, phenol/chloroform extraction. b Interference of PCR amplification by purified eluent of purification reagents. 9 μL eluent was mixed with 1 μL 166 bp of Drosophila probe DNA (0.0001 ng), and the resulting mixture was used as the template in 20 μl of real-time PCR reaction. The Ct value for Drosophila probe DNA from TE buffer was set as 100%. The experiment was repeated 3 times using de-crosslinked chromatin estimated to include 1 ng of DNA. c Size profiles of DNA purified by different reagents. The DNAs purified from de-crosslinked chromatin estimated to include 50 ng range DNA was analyzed by AATI Fragment Analyzer. DNA size (bp) is shown

    Article Snippet: The ChIP DNA Clean & Concentrator™ (Zymo Research; Zy), the Monarch® PCR & DNA Cleanup Kit (New England Biolabs; Ne), the MinElute PCR Purification Kit (Qiagen, Qm), the QIAquick PCR Purification Kit (Qiagen; Qp), the Agencourt AMPure XP kit (Beckman; Ba) and the RNAClean™ XP kit (Beckman; Br), and phenol/chloroform extraction (Invitrogen; PC) performed well with de-crosslinked chromatin.

    Techniques: DNA Purification, Chromatin Immunoprecipitation, Purification, Generated, Derivative Assay, Polymerase Chain Reaction, Amplification, Real-time Polymerase Chain Reaction

    Detection by reverse transcription–polymerase chain reaction (RT–PCR) of MUC1 transcript in activated T cells. (a) Human T cells were activated in vitro by anti-CD3 antibody. Total RNA was extracted and RT–PCR performed on the indicated days (D0, day 0; D1, day 1; D2, day 2; D3, day 3; D4, day 4). MUC1 transcript (446 bp) was identified after at least 24 hr. Human β-actin was included as a positive control, and a molecular weight marker is present in lane 1. (b) Semiquantitative RT–PCR for MUC1 in activated T cells and autologous breast cancer. Total RNA was extracted from autologous breast cancer cells and from purified human T cells following 4 days of in vitro activation using anti-CD3 antibody. cDNA was synthesized and then used as the template for RT–PCR at the dilutions shown. Human β-actin was included as a positive control.

    Journal: Immunology

    Article Title: Form and pattern of MUC1 expression on T cells activated in vivo or in vitro suggests a function in T-cell migration

    doi: 10.1046/j.1365-2567.2003.01562.x

    Figure Lengend Snippet: Detection by reverse transcription–polymerase chain reaction (RT–PCR) of MUC1 transcript in activated T cells. (a) Human T cells were activated in vitro by anti-CD3 antibody. Total RNA was extracted and RT–PCR performed on the indicated days (D0, day 0; D1, day 1; D2, day 2; D3, day 3; D4, day 4). MUC1 transcript (446 bp) was identified after at least 24 hr. Human β-actin was included as a positive control, and a molecular weight marker is present in lane 1. (b) Semiquantitative RT–PCR for MUC1 in activated T cells and autologous breast cancer. Total RNA was extracted from autologous breast cancer cells and from purified human T cells following 4 days of in vitro activation using anti-CD3 antibody. cDNA was synthesized and then used as the template for RT–PCR at the dilutions shown. Human β-actin was included as a positive control.

    Article Snippet: Total RNA was prepared using Trizol reagent (Gibco BRL) according to the manufacturer's instructions. cDNA was generated from total RNA using the reverse transcription–polymerase chain reaction (RT–PCR) kit (Stratagene, La Jolla, CA), according to the manufacturer's instructions (in all experiments 10 µg of total RNA was used to generate cDNA).

    Techniques: Reverse Transcription Polymerase Chain Reaction, In Vitro, Positive Control, Molecular Weight, Marker, Purification, Activation Assay, Synthesized