dntps  (Qiagen)


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    Structured Review

    Qiagen dntps
    Dntps, supplied by Qiagen, used in various techniques. Bioz Stars score: 96/100, based on 826 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dntps/product/Qiagen
    Average 96 stars, based on 826 article reviews
    Price from $9.99 to $1999.99
    dntps - by Bioz Stars, 2020-02
    96/100 stars

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    Related Articles

    Amplification:

    Article Title: No direct effect of the -521 C/T polymorphism in the human dopamine D4 receptor gene promoter on transcriptional activity
    Article Snippet: .. The reaction was incubated at 37°C for 40 min followed by 42°C for 20 min and 99°C for 5 min. Single-stranded cDNAs were amplified by DRD4 specific primers DART-3 (5' gca ccg cct cca tct tca acc 3') and DART-4 (5' cgg aac gtg gcc cag tag agc 3') in a final volume of 10 μl containing 1 μl of RT reaction mix, 1 μM of each primer, 5 U HotStarTaq DNA-polymerase, 1x HotStar PCR buffer, 1x Q solution and 0.2 mM dNTPs (Qiagen). .. Thermocycling conditions were as follows: incubation at 95°C for 15 min followed by 32 cycles of 94°C for 1 min, 68°C for 1 min and 72°C for 1 min completed with a final extension step at 72°C for 10 min. As described earlier, human β-actin gene served as an internal control [ ].

    Article Title: Tissue memory B cell repertoire analysis after ALVAC/AIDSVAX B/E gp120 immunization of rhesus macaques
    Article Snippet: Paragraph title: PCR amplification of immunoglobulin VH and VL genes. ... Each 50-μl reaction contained 5 μl cDNA, 0.6 μl HotStar Taq (Qiagen), 5 μl PCR Buffer (Qiagen), 10 μl Q Buffer (Qiagen), 0.4 μl 25 mM dNTPs (Qiagen), 25 mM MgCl2 (1 μl for IgH, 2 μl for Igκ and 3 μl for Igλ), and 0.125 μM IgH, Igκ, or Igλ variable region primer mixtures.

    Article Title: Structural basis for enabling T-cell receptor diversity within biased virus-specific CD8+ T-cell responses
    Article Snippet: .. For transcription of cDNA from single, sorted cells, 5 μL of cDNA mix containing 0.25 μL of Sensiscript reverse transcriptase, 1× cDNA buffer, 0.5 mM dNTPs (Qiagen), 0.125 μg of oligo(dT) (Promega), 100 μg/mL gelatin (Roche), 100 μg/mL tRNA (Roche), 20 U RNase OUT (Invitrogen Life Technologies), and 0.1% Triton X-100 (Sigma-Aldrich) was added to wells and plates incubated at 37 °C for 90 min. Plates were stored at −80 °C and the TRAV21 and TRBV29 transcripts were later amplified by nested PCR and sequenced ( ). .. Details of the conditions utlised for expression, refolding and crystalization of the 6218 TCRαβ-H2Db -PA224 complex can be found in the .

    Article Title: Excitation of Ventral Tegmental Area Dopaminergic and Nondopaminergic Neurons by Orexins/Hypocretins
    Article Snippet: The identity of cDNA sequences was revealed by sequencing the second-round amplification products, performed as described by . .. Taq enzyme, PCR buffer, Mg2+ solution, and four dNTPs were purchased from Qiagen (Erkrath, Germany).

    Article Title: Protein Phosphatase 2A Regulates bim Expression via the Akt/FKHRL1 Signaling Pathway in Amyloid-β Peptide-Induced Cerebrovascular Endothelial Cell Death
    Article Snippet: Equal amounts of total RNA (600 ng) were reverse transcribed with 1 μmol/L oligo (dT) and 0.5 mmol/L dNTPs (Qiagen), 10 U of Rnasin (Promega, Madison, WI), and 4 U of reverse transcriptase (Qiagen) for 1 h at 37°C. .. A standard amplification program was used (1 cycle of 50°C for 2 min, 1 cycle of 95°C for 10 min, 45 cycles of 95°C for 15 s, and 60°C for 1 min).

    Article Title: Host Life History Strategy, Species Diversity, and Habitat Influence Trypanosoma cruzi Vector Infection in Changing Landscapes
    Article Snippet: .. A 25 l reaction was prepared for PCR amplification with 3.0 mM MgCl2 (Fermentas), 0.2 mM dNTPs (Qiagen), 0.8 M of each primer, of Taq buffer, and 2.35 U of Taq DNA polymerase (Fermentas). ..

    Article Title: Activation of TGF-β within cultured hepatocytes and in liver injury leads to intracrine signaling with expression of connective tissue growth factor
    Article Snippet: The size of the amplified TGF-β1 fragment was 450 bp and the size of the amplified decorin fragment was 214kb. .. In general, 1 μl cDNA in 14.5 μL of RNAse free water (Roche Diagnostics, Mannheim, Germany) and 5 μl 5 × Taq Master PCR Enhancer (Eppendorf, Hamburg, Germany) was mixed with 0.5 μl dNTPs (Qiagen), 2.5 μl of 10 × PCR Buffer/15 mM MgCl2 (Qiagen), 0.5 μl of 10 μM forward primer, 0.5 μl of 10 μM reverse primer and 0.5 μl of Taq polymerase (1U/μl; Roche Diagnostics).

    Article Title: Tigecycline Resistant Klebsiella pneumoniae Isolated from Austrian River Water
    Article Snippet: Screening for Mutations The genes ramR , marR , soxR and rpsJ were amplified and sequenced with the primers described previously [ , ]. .. We used Taq DNA polymerase and dNTPs from QIAGEN (Hilden, Germany), and a T3000 Biometra thermocycler (Biometra, Germany).

    Article Title: Genetic basis for queen-worker dimorphism in a social insect
    Article Snippet: Paragraph title: Microsatellite Amplification. ... Two microliters of each sample were used in 15 μl of PCRs containing 0.5 units of Taq DNA Polymerase (Qiagen, Chatsworth, CA), 0.8 mM dNTPs, 1× Q-solution (Qiagen), 1× PCR buffer (Qiagen), and 0.5 μM of each primer, where the reverse primer was fluorescently labeled with 6-FAM (Operon).

    Synthesized:

    Article Title: Excitation of Ventral Tegmental Area Dopaminergic and Nondopaminergic Neurons by Orexins/Hypocretins
    Article Snippet: Taq enzyme, PCR buffer, Mg2+ solution, and four dNTPs were purchased from Qiagen (Erkrath, Germany). .. Oligonucleotides were synthesized by MWG Biotech (Ebersberg, Germany), and amplification was performed on a thermal cycler (GenAmp 9600; PerkinElmer Life Sciences, Weiterstadt, Germany).

    Real-time Polymerase Chain Reaction:

    Article Title: Protein Phosphatase 2A Regulates bim Expression via the Akt/FKHRL1 Signaling Pathway in Amyloid-β Peptide-Induced Cerebrovascular Endothelial Cell Death
    Article Snippet: Paragraph title: Real-time PCR and reverse transcription-PCR. ... Equal amounts of total RNA (600 ng) were reverse transcribed with 1 μmol/L oligo (dT) and 0.5 mmol/L dNTPs (Qiagen), 10 U of Rnasin (Promega, Madison, WI), and 4 U of reverse transcriptase (Qiagen) for 1 h at 37°C.

    Article Title: Decrease of the DNA methylation levels of the ADRB3 gene in leukocytes is related with serum folate in eutrophic adults
    Article Snippet: The analysis of methylation levels in genomic DNA from blood was performed by High Resolution Melting (HRM) Real Time PCR method in a applied biosystems 7500 fast system. .. PCR was performed in a total volume of 20 μl containing: 1× buffer, 4 mM Mg2+ , 200 μM of each dNTPs (Qiagen), 250 nM of each primer, 5 μM SYTO® ), F:5′-TAGGTGATTTGGGAGATTTTTTTT-3′ and R:5′-CCCCTAACAACCCACTAATATTAAC-3′.

    Incubation:

    Article Title: No direct effect of the -521 C/T polymorphism in the human dopamine D4 receptor gene promoter on transcriptional activity
    Article Snippet: .. The reaction was incubated at 37°C for 40 min followed by 42°C for 20 min and 99°C for 5 min. Single-stranded cDNAs were amplified by DRD4 specific primers DART-3 (5' gca ccg cct cca tct tca acc 3') and DART-4 (5' cgg aac gtg gcc cag tag agc 3') in a final volume of 10 μl containing 1 μl of RT reaction mix, 1 μM of each primer, 5 U HotStarTaq DNA-polymerase, 1x HotStar PCR buffer, 1x Q solution and 0.2 mM dNTPs (Qiagen). .. Thermocycling conditions were as follows: incubation at 95°C for 15 min followed by 32 cycles of 94°C for 1 min, 68°C for 1 min and 72°C for 1 min completed with a final extension step at 72°C for 10 min. As described earlier, human β-actin gene served as an internal control [ ].

    Article Title: Tissue memory B cell repertoire analysis after ALVAC/AIDSVAX B/E gp120 immunization of rhesus macaques
    Article Snippet: A 65°C, 5-minute pre-RT incubation was performed by adding 0.1 μM of a mixture of Ig constant region primers (including IgA, IgM, IgD, IgG, and IgE) to each well. .. Each 50-μl reaction contained 5 μl cDNA, 0.6 μl HotStar Taq (Qiagen), 5 μl PCR Buffer (Qiagen), 10 μl Q Buffer (Qiagen), 0.4 μl 25 mM dNTPs (Qiagen), 25 mM MgCl2 (1 μl for IgH, 2 μl for Igκ and 3 μl for Igλ), and 0.125 μM IgH, Igκ, or Igλ variable region primer mixtures.

    Article Title: Structural basis for enabling T-cell receptor diversity within biased virus-specific CD8+ T-cell responses
    Article Snippet: .. For transcription of cDNA from single, sorted cells, 5 μL of cDNA mix containing 0.25 μL of Sensiscript reverse transcriptase, 1× cDNA buffer, 0.5 mM dNTPs (Qiagen), 0.125 μg of oligo(dT) (Promega), 100 μg/mL gelatin (Roche), 100 μg/mL tRNA (Roche), 20 U RNase OUT (Invitrogen Life Technologies), and 0.1% Triton X-100 (Sigma-Aldrich) was added to wells and plates incubated at 37 °C for 90 min. Plates were stored at −80 °C and the TRAV21 and TRBV29 transcripts were later amplified by nested PCR and sequenced ( ). .. Details of the conditions utlised for expression, refolding and crystalization of the 6218 TCRαβ-H2Db -PA224 complex can be found in the .

    Article Title: Tigecycline Resistant Klebsiella pneumoniae Isolated from Austrian River Water
    Article Snippet: Standard PCR protocols and conditions were used in the following way: initial denaturation at 94 °C for 5 min; 35 cycles of 94 °C for 30 s, 52 °C for 1 min and 72 °C for 1 min; and a final incubation for 5 min at 72 °C. .. We used Taq DNA polymerase and dNTPs from QIAGEN (Hilden, Germany), and a T3000 Biometra thermocycler (Biometra, Germany).

    Infection:

    Article Title: Structural basis for enabling T-cell receptor diversity within biased virus-specific CD8+ T-cell responses
    Article Snippet: Splenocytes were isolated from either naive mice or from immune mice at 10 d after infection and stained in sort buffer (0.1% BSA in PBS) with various combinations of Db PA224 -tetramer, anti–CD8α-PerCPCy5.5, anti–Vβ7-PE, and biotinylated anti-CD44 (detected following incubation with Streptavidin APC-Cy7) as above. .. For transcription of cDNA from single, sorted cells, 5 μL of cDNA mix containing 0.25 μL of Sensiscript reverse transcriptase, 1× cDNA buffer, 0.5 mM dNTPs (Qiagen), 0.125 μg of oligo(dT) (Promega), 100 μg/mL gelatin (Roche), 100 μg/mL tRNA (Roche), 20 U RNase OUT (Invitrogen Life Technologies), and 0.1% Triton X-100 (Sigma-Aldrich) was added to wells and plates incubated at 37 °C for 90 min. Plates were stored at −80 °C and the TRAV21 and TRBV29 transcripts were later amplified by nested PCR and sequenced ( ).

    Expressing:

    Article Title: Enhancement of Enteric Adenovirus Cultivation by Viral Transactivator Proteins ▿
    Article Snippet: To check CMV IE1 mRNA expression, we extracted mRNA using an RNeasy Mini kit (Qiagen) from the cell pellets described above. .. The RT reaction mixture contained 2 μl of 10× Buffer RT (Qiagen), 2 μl of the dNTPs (5 mM; Qiagen), 2 μl oligo(dT) primer (10 pmol/μl), 1 μl of RNase inhibitor (10 units/μl; TaKaRa, Shiga, Japan), 1 μl of Omniscript reverse transcriptase (Qiagen), and 5 μl of mRNA template in a final volume of 20 μl.

    Article Title: Protein Phosphatase 2A Regulates bim Expression via the Akt/FKHRL1 Signaling Pathway in Amyloid-β Peptide-Induced Cerebrovascular Endothelial Cell Death
    Article Snippet: Real-time PCR was used to measure mRNA expression of BH3-only proteins. .. Equal amounts of total RNA (600 ng) were reverse transcribed with 1 μmol/L oligo (dT) and 0.5 mmol/L dNTPs (Qiagen), 10 U of Rnasin (Promega, Madison, WI), and 4 U of reverse transcriptase (Qiagen) for 1 h at 37°C.

    Transformation Assay:

    Article Title: Decrease of the DNA methylation levels of the ADRB3 gene in leukocytes is related with serum folate in eutrophic adults
    Article Snippet: The leukocyte DNA was isolated, quantified and transformed with sodium bisulfite according to the conditions described in a previously published study [ ]. .. PCR was performed in a total volume of 20 μl containing: 1× buffer, 4 mM Mg2+ , 200 μM of each dNTPs (Qiagen), 250 nM of each primer, 5 μM SYTO® ), F:5′-TAGGTGATTTGGGAGATTTTTTTT-3′ and R:5′-CCCCTAACAACCCACTAATATTAAC-3′.

    Countercurrent Chromatography:

    Article Title: Enhancement of Enteric Adenovirus Cultivation by Viral Transactivator Proteins ▿
    Article Snippet: A forward (5′-GAT CCC CTC GCG AGT TGG TTC-3′) and reverse (5′-AAT GGG GCG GAG TTG TTA CGA C-3′) primer set was designed based on the pLNCX vector that would amplify a product of 834 bp. .. The RT reaction mixture contained 2 μl of 10× Buffer RT (Qiagen), 2 μl of the dNTPs (5 mM; Qiagen), 2 μl oligo(dT) primer (10 pmol/μl), 1 μl of RNase inhibitor (10 units/μl; TaKaRa, Shiga, Japan), 1 μl of Omniscript reverse transcriptase (Qiagen), and 5 μl of mRNA template in a final volume of 20 μl.

    Article Title: Activation of TGF-β within cultured hepatocytes and in liver injury leads to intracrine signaling with expression of connective tissue growth factor
    Article Snippet: RT-PCR was performed using the Biometra T3000 Thermocycler PCR System (Biometra, Göttingen, Germany) and the following primers: rat TGF-β1 (forward: 5′-CTC TCC ACC TGC AAG AC-3′; reverse: 5′-GGA CTC TCC ACC TGC AAG AC-3′) and rat decorin (forward: 5′-CTC TGG CAT AAT CCC TTA CGA-3′; reverse: 5′-GGT ATG CAA GTC CTT CAG GTT-3′). .. In general, 1 μl cDNA in 14.5 μL of RNAse free water (Roche Diagnostics, Mannheim, Germany) and 5 μl 5 × Taq Master PCR Enhancer (Eppendorf, Hamburg, Germany) was mixed with 0.5 μl dNTPs (Qiagen), 2.5 μl of 10 × PCR Buffer/15 mM MgCl2 (Qiagen), 0.5 μl of 10 μM forward primer, 0.5 μl of 10 μM reverse primer and 0.5 μl of Taq polymerase (1U/μl; Roche Diagnostics).

    Electroporation:

    Article Title: Enhancement of Enteric Adenovirus Cultivation by Viral Transactivator Proteins ▿
    Article Snippet: Paragraph title: Construction of the new cell line 293-CMV by electroporation. ... The RT reaction mixture contained 2 μl of 10× Buffer RT (Qiagen), 2 μl of the dNTPs (5 mM; Qiagen), 2 μl oligo(dT) primer (10 pmol/μl), 1 μl of RNase inhibitor (10 units/μl; TaKaRa, Shiga, Japan), 1 μl of Omniscript reverse transcriptase (Qiagen), and 5 μl of mRNA template in a final volume of 20 μl.

    Cell Culture:

    Article Title: Protein Phosphatase 2A Regulates bim Expression via the Akt/FKHRL1 Signaling Pathway in Amyloid-β Peptide-Induced Cerebrovascular Endothelial Cell Death
    Article Snippet: Total RNA from the mouse brain microvessel fraction or cultured mouse CECs were isolated with the RNeasy Mini kit (Qiagen, Valencia, CA). .. Equal amounts of total RNA (600 ng) were reverse transcribed with 1 μmol/L oligo (dT) and 0.5 mmol/L dNTPs (Qiagen), 10 U of Rnasin (Promega, Madison, WI), and 4 U of reverse transcriptase (Qiagen) for 1 h at 37°C.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: No direct effect of the -521 C/T polymorphism in the human dopamine D4 receptor gene promoter on transcriptional activity
    Article Snippet: Paragraph title: Total RNA isolation, RT-PCR ... The reaction was incubated at 37°C for 40 min followed by 42°C for 20 min and 99°C for 5 min. Single-stranded cDNAs were amplified by DRD4 specific primers DART-3 (5' gca ccg cct cca tct tca acc 3') and DART-4 (5' cgg aac gtg gcc cag tag agc 3') in a final volume of 10 μl containing 1 μl of RT reaction mix, 1 μM of each primer, 5 U HotStarTaq DNA-polymerase, 1x HotStar PCR buffer, 1x Q solution and 0.2 mM dNTPs (Qiagen).

    Article Title: Tissue memory B cell repertoire analysis after ALVAC/AIDSVAX B/E gp120 immunization of rhesus macaques
    Article Snippet: First round PCR (PCRa) of heavy, kappa, and lambda chains was then performed using the cDNA product of RT/PCR. .. Each 50-μl reaction contained 5 μl cDNA, 0.6 μl HotStar Taq (Qiagen), 5 μl PCR Buffer (Qiagen), 10 μl Q Buffer (Qiagen), 0.4 μl 25 mM dNTPs (Qiagen), 25 mM MgCl2 (1 μl for IgH, 2 μl for Igκ and 3 μl for Igλ), and 0.125 μM IgH, Igκ, or Igλ variable region primer mixtures.

    Article Title: Structural basis for enabling T-cell receptor diversity within biased virus-specific CD8+ T-cell responses
    Article Snippet: Paragraph title: Isolation of CD8+ T Cells, RT-PCR, and Sequencing. ... For transcription of cDNA from single, sorted cells, 5 μL of cDNA mix containing 0.25 μL of Sensiscript reverse transcriptase, 1× cDNA buffer, 0.5 mM dNTPs (Qiagen), 0.125 μg of oligo(dT) (Promega), 100 μg/mL gelatin (Roche), 100 μg/mL tRNA (Roche), 20 U RNase OUT (Invitrogen Life Technologies), and 0.1% Triton X-100 (Sigma-Aldrich) was added to wells and plates incubated at 37 °C for 90 min. Plates were stored at −80 °C and the TRAV21 and TRBV29 transcripts were later amplified by nested PCR and sequenced ( ).

    Article Title: Activation of TGF-β within cultured hepatocytes and in liver injury leads to intracrine signaling with expression of connective tissue growth factor
    Article Snippet: Paragraph title: RT-PCR for TGF-β1 ... In general, 1 μl cDNA in 14.5 μL of RNAse free water (Roche Diagnostics, Mannheim, Germany) and 5 μl 5 × Taq Master PCR Enhancer (Eppendorf, Hamburg, Germany) was mixed with 0.5 μl dNTPs (Qiagen), 2.5 μl of 10 × PCR Buffer/15 mM MgCl2 (Qiagen), 0.5 μl of 10 μM forward primer, 0.5 μl of 10 μM reverse primer and 0.5 μl of Taq polymerase (1U/μl; Roche Diagnostics).

    Sequencing:

    Article Title: Structural basis for enabling T-cell receptor diversity within biased virus-specific CD8+ T-cell responses
    Article Snippet: Paragraph title: Isolation of CD8+ T Cells, RT-PCR, and Sequencing. ... For transcription of cDNA from single, sorted cells, 5 μL of cDNA mix containing 0.25 μL of Sensiscript reverse transcriptase, 1× cDNA buffer, 0.5 mM dNTPs (Qiagen), 0.125 μg of oligo(dT) (Promega), 100 μg/mL gelatin (Roche), 100 μg/mL tRNA (Roche), 20 U RNase OUT (Invitrogen Life Technologies), and 0.1% Triton X-100 (Sigma-Aldrich) was added to wells and plates incubated at 37 °C for 90 min. Plates were stored at −80 °C and the TRAV21 and TRBV29 transcripts were later amplified by nested PCR and sequenced ( ).

    Article Title: Excitation of Ventral Tegmental Area Dopaminergic and Nondopaminergic Neurons by Orexins/Hypocretins
    Article Snippet: The identity of cDNA sequences was revealed by sequencing the second-round amplification products, performed as described by . .. Taq enzyme, PCR buffer, Mg2+ solution, and four dNTPs were purchased from Qiagen (Erkrath, Germany).

    Article Title: Protein Phosphatase 2A Regulates bim Expression via the Akt/FKHRL1 Signaling Pathway in Amyloid-β Peptide-Induced Cerebrovascular Endothelial Cell Death
    Article Snippet: Equal amounts of total RNA (600 ng) were reverse transcribed with 1 μmol/L oligo (dT) and 0.5 mmol/L dNTPs (Qiagen), 10 U of Rnasin (Promega, Madison, WI), and 4 U of reverse transcriptase (Qiagen) for 1 h at 37°C. .. Ten nanograms of cDNA were subjected to PCR using the ABI Prism 7000 Sequence Detection System and SYBR Green PCR Master Mix (both from Applied Biosystems, Foster City, CA).

    Article Title: Tigecycline Resistant Klebsiella pneumoniae Isolated from Austrian River Water
    Article Snippet: We used Taq DNA polymerase and dNTPs from QIAGEN (Hilden, Germany), and a T3000 Biometra thermocycler (Biometra, Germany). .. Sequencing was performed with the Mix2Seq Kit (Eurofins Genomics).

    Cellular Antioxidant Activity Assay:

    Article Title: Activation of TGF-β within cultured hepatocytes and in liver injury leads to intracrine signaling with expression of connective tissue growth factor
    Article Snippet: RT-PCR was performed using the Biometra T3000 Thermocycler PCR System (Biometra, Göttingen, Germany) and the following primers: rat TGF-β1 (forward: 5′-CTC TCC ACC TGC AAG AC-3′; reverse: 5′-GGA CTC TCC ACC TGC AAG AC-3′) and rat decorin (forward: 5′-CTC TGG CAT AAT CCC TTA CGA-3′; reverse: 5′-GGT ATG CAA GTC CTT CAG GTT-3′). .. In general, 1 μl cDNA in 14.5 μL of RNAse free water (Roche Diagnostics, Mannheim, Germany) and 5 μl 5 × Taq Master PCR Enhancer (Eppendorf, Hamburg, Germany) was mixed with 0.5 μl dNTPs (Qiagen), 2.5 μl of 10 × PCR Buffer/15 mM MgCl2 (Qiagen), 0.5 μl of 10 μM forward primer, 0.5 μl of 10 μM reverse primer and 0.5 μl of Taq polymerase (1U/μl; Roche Diagnostics).

    DNA Extraction:

    Article Title: Specificity of the Effect of a Nicotinic Receptor Polymorphism on Individual Differences in Visuospatial Attention
    Article Snippet: Buccal (cheek) swabs were obtained from each participant in order to extract DNA ( ) and prepared as directed by the manufacturer (MasterAMP TMBuccal Swab DNA Extraction Kit, Epicentre Technologies, Madison, WI). .. Taq polymerase, PCR buffer, and dNTPs were obtained from QIAGEN (Valencia, CA) and used at recommended concentrations for a 20-μl PCR reaction.

    Nucleic Acid Electrophoresis:

    Article Title: Aboveground Whitefly Infestation Modulates Transcriptional Levels of Anthocyanin Biosynthesis and Jasmonic Acid Signaling-Related Genes and Augments the Cope with Drought Stress of Maize
    Article Snippet: RNA quality and quantity were confirmed by gel electrophoresis and a Nanodrop spectrophotometer (ND-1000, DE, USA) before use in subsequent analyses. .. Synthesis of first-strand cDNA was performed from 1 μg of total RNA as a template with oligo-dT primer (Qiagen, CA, USA), dNTPs (Qiagen, CA, USA), and Moloney murine leukemia virus reverse transcriptase (MMLV-RT; Enzynomics, Daejeon, South Korea).

    Article Title: Specificity of the Effect of a Nicotinic Receptor Polymorphism on Individual Differences in Visuospatial Attention
    Article Snippet: Taq polymerase, PCR buffer, and dNTPs were obtained from QIAGEN (Valencia, CA) and used at recommended concentrations for a 20-μl PCR reaction. .. Gel electrophoresis in either LE or Metaphor agarose, followed by staining in ethidium bromide, was used to resolve and visualize DNA fragments.

    Fluorescence:

    Article Title: Decrease of the DNA methylation levels of the ADRB3 gene in leukocytes is related with serum folate in eutrophic adults
    Article Snippet: PCR was performed in a total volume of 20 μl containing: 1× buffer, 4 mM Mg2+ , 200 μM of each dNTPs (Qiagen), 250 nM of each primer, 5 μM SYTO® ), F:5′-TAGGTGATTTGGGAGATTTTTTTT-3′ and R:5′-CCCCTAACAACCCACTAATATTAAC-3′. .. The melting curves were normalized by calculation of the ‘line of best fit’ in between two normalization regions before and after the major fluorescence decrease representing the melting of the PCR product using the software provided with the HRM Software v2.0, provided by 7500 fast system.

    Methylation:

    Article Title: Decrease of the DNA methylation levels of the ADRB3 gene in leukocytes is related with serum folate in eutrophic adults
    Article Snippet: The analysis of methylation levels in genomic DNA from blood was performed by High Resolution Melting (HRM) Real Time PCR method in a applied biosystems 7500 fast system. .. PCR was performed in a total volume of 20 μl containing: 1× buffer, 4 mM Mg2+ , 200 μM of each dNTPs (Qiagen), 250 nM of each primer, 5 μM SYTO® ), F:5′-TAGGTGATTTGGGAGATTTTTTTT-3′ and R:5′-CCCCTAACAACCCACTAATATTAAC-3′.

    Isolation:

    Article Title: No direct effect of the -521 C/T polymorphism in the human dopamine D4 receptor gene promoter on transcriptional activity
    Article Snippet: Paragraph title: Total RNA isolation, RT-PCR ... The reaction was incubated at 37°C for 40 min followed by 42°C for 20 min and 99°C for 5 min. Single-stranded cDNAs were amplified by DRD4 specific primers DART-3 (5' gca ccg cct cca tct tca acc 3') and DART-4 (5' cgg aac gtg gcc cag tag agc 3') in a final volume of 10 μl containing 1 μl of RT reaction mix, 1 μM of each primer, 5 U HotStarTaq DNA-polymerase, 1x HotStar PCR buffer, 1x Q solution and 0.2 mM dNTPs (Qiagen).

    Article Title: Structural basis for enabling T-cell receptor diversity within biased virus-specific CD8+ T-cell responses
    Article Snippet: Paragraph title: Isolation of CD8+ T Cells, RT-PCR, and Sequencing. ... For transcription of cDNA from single, sorted cells, 5 μL of cDNA mix containing 0.25 μL of Sensiscript reverse transcriptase, 1× cDNA buffer, 0.5 mM dNTPs (Qiagen), 0.125 μg of oligo(dT) (Promega), 100 μg/mL gelatin (Roche), 100 μg/mL tRNA (Roche), 20 U RNase OUT (Invitrogen Life Technologies), and 0.1% Triton X-100 (Sigma-Aldrich) was added to wells and plates incubated at 37 °C for 90 min. Plates were stored at −80 °C and the TRAV21 and TRBV29 transcripts were later amplified by nested PCR and sequenced ( ).

    Article Title: Aboveground Whitefly Infestation Modulates Transcriptional Levels of Anthocyanin Biosynthesis and Jasmonic Acid Signaling-Related Genes and Augments the Cope with Drought Stress of Maize
    Article Snippet: Total RNA was isolated from ground tissues using a previously published method [ ]. .. Synthesis of first-strand cDNA was performed from 1 μg of total RNA as a template with oligo-dT primer (Qiagen, CA, USA), dNTPs (Qiagen, CA, USA), and Moloney murine leukemia virus reverse transcriptase (MMLV-RT; Enzynomics, Daejeon, South Korea).

    Article Title: Protein Phosphatase 2A Regulates bim Expression via the Akt/FKHRL1 Signaling Pathway in Amyloid-β Peptide-Induced Cerebrovascular Endothelial Cell Death
    Article Snippet: Total RNA from the mouse brain microvessel fraction or cultured mouse CECs were isolated with the RNeasy Mini kit (Qiagen, Valencia, CA). .. Equal amounts of total RNA (600 ng) were reverse transcribed with 1 μmol/L oligo (dT) and 0.5 mmol/L dNTPs (Qiagen), 10 U of Rnasin (Promega, Madison, WI), and 4 U of reverse transcriptase (Qiagen) for 1 h at 37°C.

    Size-exclusion Chromatography:

    Article Title: Excitation of Ventral Tegmental Area Dopaminergic and Nondopaminergic Neurons by Orexins/Hypocretins
    Article Snippet: Taq enzyme, PCR buffer, Mg2+ solution, and four dNTPs were purchased from Qiagen (Erkrath, Germany). .. In each round, 35 cycles of the following thermal programs were used: denaturation at 94°C for 48 sec, annealing at 53°C for 1 min, and extension at 72°C for 90 sec. For the second amplification round, 1 μl of product from the first amplification was used as a template.

    Labeling:

    Article Title: Genetic basis for queen-worker dimorphism in a social insect
    Article Snippet: .. Two microliters of each sample were used in 15 μl of PCRs containing 0.5 units of Taq DNA Polymerase (Qiagen, Chatsworth, CA), 0.8 mM dNTPs, 1× Q-solution (Qiagen), 1× PCR buffer (Qiagen), and 0.5 μM of each primer, where the reverse primer was fluorescently labeled with 6-FAM (Operon). .. The PCR cycle consisted of 95°C for 5 min, followed by 25 cycles of 95°C for 30 s, 48°C for 30 s, 72°C for 30 s, followed by 72°C for 7 min.

    Mouse Assay:

    Article Title: Structural basis for enabling T-cell receptor diversity within biased virus-specific CD8+ T-cell responses
    Article Snippet: Splenocytes were isolated from either naive mice or from immune mice at 10 d after infection and stained in sort buffer (0.1% BSA in PBS) with various combinations of Db PA224 -tetramer, anti–CD8α-PerCPCy5.5, anti–Vβ7-PE, and biotinylated anti-CD44 (detected following incubation with Streptavidin APC-Cy7) as above. .. For transcription of cDNA from single, sorted cells, 5 μL of cDNA mix containing 0.25 μL of Sensiscript reverse transcriptase, 1× cDNA buffer, 0.5 mM dNTPs (Qiagen), 0.125 μg of oligo(dT) (Promega), 100 μg/mL gelatin (Roche), 100 μg/mL tRNA (Roche), 20 U RNase OUT (Invitrogen Life Technologies), and 0.1% Triton X-100 (Sigma-Aldrich) was added to wells and plates incubated at 37 °C for 90 min. Plates were stored at −80 °C and the TRAV21 and TRBV29 transcripts were later amplified by nested PCR and sequenced ( ).

    Polymerase Chain Reaction:

    Article Title: No direct effect of the -521 C/T polymorphism in the human dopamine D4 receptor gene promoter on transcriptional activity
    Article Snippet: .. The reaction was incubated at 37°C for 40 min followed by 42°C for 20 min and 99°C for 5 min. Single-stranded cDNAs were amplified by DRD4 specific primers DART-3 (5' gca ccg cct cca tct tca acc 3') and DART-4 (5' cgg aac gtg gcc cag tag agc 3') in a final volume of 10 μl containing 1 μl of RT reaction mix, 1 μM of each primer, 5 U HotStarTaq DNA-polymerase, 1x HotStar PCR buffer, 1x Q solution and 0.2 mM dNTPs (Qiagen). .. Thermocycling conditions were as follows: incubation at 95°C for 15 min followed by 32 cycles of 94°C for 1 min, 68°C for 1 min and 72°C for 1 min completed with a final extension step at 72°C for 10 min. As described earlier, human β-actin gene served as an internal control [ ].

    Article Title: Enhancement of Enteric Adenovirus Cultivation by Viral Transactivator Proteins ▿
    Article Snippet: The PCR assay was performed under the following conditions: denaturation for 2 min at 94°C and then 40 cycles of denaturation for 30 s at 94°C, annealing for 30 s at 55°C, and extension for 30 s at 68°C. .. The RT reaction mixture contained 2 μl of 10× Buffer RT (Qiagen), 2 μl of the dNTPs (5 mM; Qiagen), 2 μl oligo(dT) primer (10 pmol/μl), 1 μl of RNase inhibitor (10 units/μl; TaKaRa, Shiga, Japan), 1 μl of Omniscript reverse transcriptase (Qiagen), and 5 μl of mRNA template in a final volume of 20 μl.

    Article Title: Tissue memory B cell repertoire analysis after ALVAC/AIDSVAX B/E gp120 immunization of rhesus macaques
    Article Snippet: .. Each 50-μl reaction contained 5 μl cDNA, 0.6 μl HotStar Taq (Qiagen), 5 μl PCR Buffer (Qiagen), 10 μl Q Buffer (Qiagen), 0.4 μl 25 mM dNTPs (Qiagen), 25 mM MgCl2 (1 μl for IgH, 2 μl for Igκ and 3 μl for Igλ), and 0.125 μM IgH, Igκ, or Igλ variable region primer mixtures. .. PCRa reaction conditions were as follows: 95°C/5 minutes, (94°C/30 seconds, 62°C [for IgH] or 64°C [for Igκ/λ]/45 seconds, 72°C/90 seconds) for 35 cycles, 72°C/7 minutes, then hold at 10°C.

    Article Title: Excitation of Ventral Tegmental Area Dopaminergic and Nondopaminergic Neurons by Orexins/Hypocretins
    Article Snippet: .. Taq enzyme, PCR buffer, Mg2+ solution, and four dNTPs were purchased from Qiagen (Erkrath, Germany). .. Oligonucleotides were synthesized by MWG Biotech (Ebersberg, Germany), and amplification was performed on a thermal cycler (GenAmp 9600; PerkinElmer Life Sciences, Weiterstadt, Germany).

    Article Title: Specificity of the Effect of a Nicotinic Receptor Polymorphism on Individual Differences in Visuospatial Attention
    Article Snippet: .. Taq polymerase, PCR buffer, and dNTPs were obtained from QIAGEN (Valencia, CA) and used at recommended concentrations for a 20-μl PCR reaction. .. PCR reactions and restriction digests (PCR-RFLP) were performed on the PTC-100 Programmable Thermal Controller (MJ Research, Watertown, MA).

    Article Title: A synonymous germline variant in a gene encoding a cell adhesion molecule is associated with cutaneous mast cell tumour development in Labrador and Golden Retrievers
    Article Snippet: .. A 1μl aliquot of a 1 : 100 dilution of each first round PCR product was used in a 20μl second round PCR reaction comprising 1 x HotStar Taq PCR buffer (Qiagen), 1 x Q-Solution (Qiagen), 0.5μM of both the ‘internal’ (B) forward and reverse primers , 0.3mM of each of 4 x dNTPs (Qiagen), 2 units of HotStar Taq DNA Polymerase (Qiagen), and 0.08 units of HotStar HiFidelity Taq DNA Polymerase (Qiagen). .. Second round PCR reaction products were analysed by 2% agarose gel electrophoresis (E14-15 Assay) and by 0.8% agarose gel electrophoresis (E15-17 Assay), respectively.

    Article Title: Growth of White Matter in the Adolescent Brain: Role of Testosterone and Androgen Receptor
    Article Snippet: .. PCRs were performed using 100 ng of genomic DNA in a total volume of 8.0 μl containing 1.0 m m MgCl2 (Qiagen), 1× PCR buffer containing 1.5 m m MgCl2 (Qiagen), 0.035 μ m dNTPs (Qiagen), 0.04 U/μl HotstarTaq DNA polymerase (Qiagen), and 200 n m forward and reverse primer. .. PCR protocol was initiated by denaturing the samples at 95°C for 10 min followed by 45 cycles containing a denaturation phase at 95°C for 30 s, an annealing phase at 60°C for 30 s, and an extension phase at 72°C for 30 s. The final extension was done at 72°C for 7 min. A reading mixture was prepared with 2 μl of PCR products, 0.15 μl of Genescan 500 Liz size standard (Applied Biosystems), and 8.5 μl of Hi-Di Formamide (Applied Biosystems) and migrated on Applied Biosystems 3730xl DNA Analyzer.

    Article Title: Protein Phosphatase 2A Regulates bim Expression via the Akt/FKHRL1 Signaling Pathway in Amyloid-β Peptide-Induced Cerebrovascular Endothelial Cell Death
    Article Snippet: Equal amounts of total RNA (600 ng) were reverse transcribed with 1 μmol/L oligo (dT) and 0.5 mmol/L dNTPs (Qiagen), 10 U of Rnasin (Promega, Madison, WI), and 4 U of reverse transcriptase (Qiagen) for 1 h at 37°C. .. Ten nanograms of cDNA were subjected to PCR using the ABI Prism 7000 Sequence Detection System and SYBR Green PCR Master Mix (both from Applied Biosystems, Foster City, CA).

    Article Title: Host Life History Strategy, Species Diversity, and Habitat Influence Trypanosoma cruzi Vector Infection in Changing Landscapes
    Article Snippet: .. A 25 l reaction was prepared for PCR amplification with 3.0 mM MgCl2 (Fermentas), 0.2 mM dNTPs (Qiagen), 0.8 M of each primer, of Taq buffer, and 2.35 U of Taq DNA polymerase (Fermentas). ..

    Article Title: Activation of TGF-β within cultured hepatocytes and in liver injury leads to intracrine signaling with expression of connective tissue growth factor
    Article Snippet: .. In general, 1 μl cDNA in 14.5 μL of RNAse free water (Roche Diagnostics, Mannheim, Germany) and 5 μl 5 × Taq Master PCR Enhancer (Eppendorf, Hamburg, Germany) was mixed with 0.5 μl dNTPs (Qiagen), 2.5 μl of 10 × PCR Buffer/15 mM MgCl2 (Qiagen), 0.5 μl of 10 μM forward primer, 0.5 μl of 10 μM reverse primer and 0.5 μl of Taq polymerase (1U/μl; Roche Diagnostics). ..

    Article Title: Tigecycline Resistant Klebsiella pneumoniae Isolated from Austrian River Water
    Article Snippet: Standard PCR protocols and conditions were used in the following way: initial denaturation at 94 °C for 5 min; 35 cycles of 94 °C for 30 s, 52 °C for 1 min and 72 °C for 1 min; and a final incubation for 5 min at 72 °C. .. We used Taq DNA polymerase and dNTPs from QIAGEN (Hilden, Germany), and a T3000 Biometra thermocycler (Biometra, Germany).

    Article Title: Genetic basis for queen-worker dimorphism in a social insect
    Article Snippet: .. Two microliters of each sample were used in 15 μl of PCRs containing 0.5 units of Taq DNA Polymerase (Qiagen, Chatsworth, CA), 0.8 mM dNTPs, 1× Q-solution (Qiagen), 1× PCR buffer (Qiagen), and 0.5 μM of each primer, where the reverse primer was fluorescently labeled with 6-FAM (Operon). .. The PCR cycle consisted of 95°C for 5 min, followed by 25 cycles of 95°C for 30 s, 48°C for 30 s, 72°C for 30 s, followed by 72°C for 7 min.

    Article Title: Decrease of the DNA methylation levels of the ADRB3 gene in leukocytes is related with serum folate in eutrophic adults
    Article Snippet: .. PCR was performed in a total volume of 20 μl containing: 1× buffer, 4 mM Mg2+ , 200 μM of each dNTPs (Qiagen), 250 nM of each primer, 5 μM SYTO® ), F:5′-TAGGTGATTTGGGAGATTTTTTTT-3′ and R:5′-CCCCTAACAACCCACTAATATTAAC-3′. .. The PCR program consisted of an initial enzymatic activation at 95 °C for 10 min, followed by 50 cycles of 45 s at 95 °C, 45 s at 60 °C and 45 s at 72 °C and the final extension at 72 °C for 10 min.

    Staining:

    Article Title: Structural basis for enabling T-cell receptor diversity within biased virus-specific CD8+ T-cell responses
    Article Snippet: Splenocytes were isolated from either naive mice or from immune mice at 10 d after infection and stained in sort buffer (0.1% BSA in PBS) with various combinations of Db PA224 -tetramer, anti–CD8α-PerCPCy5.5, anti–Vβ7-PE, and biotinylated anti-CD44 (detected following incubation with Streptavidin APC-Cy7) as above. .. For transcription of cDNA from single, sorted cells, 5 μL of cDNA mix containing 0.25 μL of Sensiscript reverse transcriptase, 1× cDNA buffer, 0.5 mM dNTPs (Qiagen), 0.125 μg of oligo(dT) (Promega), 100 μg/mL gelatin (Roche), 100 μg/mL tRNA (Roche), 20 U RNase OUT (Invitrogen Life Technologies), and 0.1% Triton X-100 (Sigma-Aldrich) was added to wells and plates incubated at 37 °C for 90 min. Plates were stored at −80 °C and the TRAV21 and TRBV29 transcripts were later amplified by nested PCR and sequenced ( ).

    Article Title: Specificity of the Effect of a Nicotinic Receptor Polymorphism on Individual Differences in Visuospatial Attention
    Article Snippet: Taq polymerase, PCR buffer, and dNTPs were obtained from QIAGEN (Valencia, CA) and used at recommended concentrations for a 20-μl PCR reaction. .. Gel electrophoresis in either LE or Metaphor agarose, followed by staining in ethidium bromide, was used to resolve and visualize DNA fragments.

    Nested PCR:

    Article Title: Tissue memory B cell repertoire analysis after ALVAC/AIDSVAX B/E gp120 immunization of rhesus macaques
    Article Snippet: Each 50-μl reaction contained 5 μl cDNA, 0.6 μl HotStar Taq (Qiagen), 5 μl PCR Buffer (Qiagen), 10 μl Q Buffer (Qiagen), 0.4 μl 25 mM dNTPs (Qiagen), 25 mM MgCl2 (1 μl for IgH, 2 μl for Igκ and 3 μl for Igλ), and 0.125 μM IgH, Igκ, or Igλ variable region primer mixtures. .. PCRa products were used as templates for nested PCR (PCRb).

    Article Title: Structural basis for enabling T-cell receptor diversity within biased virus-specific CD8+ T-cell responses
    Article Snippet: .. For transcription of cDNA from single, sorted cells, 5 μL of cDNA mix containing 0.25 μL of Sensiscript reverse transcriptase, 1× cDNA buffer, 0.5 mM dNTPs (Qiagen), 0.125 μg of oligo(dT) (Promega), 100 μg/mL gelatin (Roche), 100 μg/mL tRNA (Roche), 20 U RNase OUT (Invitrogen Life Technologies), and 0.1% Triton X-100 (Sigma-Aldrich) was added to wells and plates incubated at 37 °C for 90 min. Plates were stored at −80 °C and the TRAV21 and TRBV29 transcripts were later amplified by nested PCR and sequenced ( ). .. Details of the conditions utlised for expression, refolding and crystalization of the 6218 TCRαβ-H2Db -PA224 complex can be found in the .

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Activation of TGF-β within cultured hepatocytes and in liver injury leads to intracrine signaling with expression of connective tissue growth factor
    Article Snippet: RT-PCR was performed using the Biometra T3000 Thermocycler PCR System (Biometra, Göttingen, Germany) and the following primers: rat TGF-β1 (forward: 5′-CTC TCC ACC TGC AAG AC-3′; reverse: 5′-GGA CTC TCC ACC TGC AAG AC-3′) and rat decorin (forward: 5′-CTC TGG CAT AAT CCC TTA CGA-3′; reverse: 5′-GGT ATG CAA GTC CTT CAG GTT-3′). .. In general, 1 μl cDNA in 14.5 μL of RNAse free water (Roche Diagnostics, Mannheim, Germany) and 5 μl 5 × Taq Master PCR Enhancer (Eppendorf, Hamburg, Germany) was mixed with 0.5 μl dNTPs (Qiagen), 2.5 μl of 10 × PCR Buffer/15 mM MgCl2 (Qiagen), 0.5 μl of 10 μM forward primer, 0.5 μl of 10 μM reverse primer and 0.5 μl of Taq polymerase (1U/μl; Roche Diagnostics).

    Purification:

    Article Title: Activation of TGF-β within cultured hepatocytes and in liver injury leads to intracrine signaling with expression of connective tissue growth factor
    Article Snippet: RT-PCR for TGF-β1 Total cellular RNA was extracted from PC and HSC, respectively, with the Qiagen RNeasy purification kit (Qiagen, Hilden, Germany). cDNA was reverse-transcribed using the First-Strand cDNA synthesis kit (Invitrogen). .. In general, 1 μl cDNA in 14.5 μL of RNAse free water (Roche Diagnostics, Mannheim, Germany) and 5 μl 5 × Taq Master PCR Enhancer (Eppendorf, Hamburg, Germany) was mixed with 0.5 μl dNTPs (Qiagen), 2.5 μl of 10 × PCR Buffer/15 mM MgCl2 (Qiagen), 0.5 μl of 10 μM forward primer, 0.5 μl of 10 μM reverse primer and 0.5 μl of Taq polymerase (1U/μl; Roche Diagnostics).

    Plasmid Preparation:

    Article Title: Enhancement of Enteric Adenovirus Cultivation by Viral Transactivator Proteins ▿
    Article Snippet: A forward (5′-GAT CCC CTC GCG AGT TGG TTC-3′) and reverse (5′-AAT GGG GCG GAG TTG TTA CGA C-3′) primer set was designed based on the pLNCX vector that would amplify a product of 834 bp. .. The RT reaction mixture contained 2 μl of 10× Buffer RT (Qiagen), 2 μl of the dNTPs (5 mM; Qiagen), 2 μl oligo(dT) primer (10 pmol/μl), 1 μl of RNase inhibitor (10 units/μl; TaKaRa, Shiga, Japan), 1 μl of Omniscript reverse transcriptase (Qiagen), and 5 μl of mRNA template in a final volume of 20 μl.

    Article Title: Host Life History Strategy, Species Diversity, and Habitat Influence Trypanosoma cruzi Vector Infection in Changing Landscapes
    Article Snippet: Due to positive template bias in the PCR reaction, it is unlikely that this assay would result in the detection of multiple blood meal sources in a single vector. .. A 25 l reaction was prepared for PCR amplification with 3.0 mM MgCl2 (Fermentas), 0.2 mM dNTPs (Qiagen), 0.8 M of each primer, of Taq buffer, and 2.35 U of Taq DNA polymerase (Fermentas).

    Software:

    Article Title: Protein Phosphatase 2A Regulates bim Expression via the Akt/FKHRL1 Signaling Pathway in Amyloid-β Peptide-Induced Cerebrovascular Endothelial Cell Death
    Article Snippet: Equal amounts of total RNA (600 ng) were reverse transcribed with 1 μmol/L oligo (dT) and 0.5 mmol/L dNTPs (Qiagen), 10 U of Rnasin (Promega, Madison, WI), and 4 U of reverse transcriptase (Qiagen) for 1 h at 37°C. .. The primers of the BH3-only genes were designed using Primer Express software (Applied Biosystems) as follows: bim (forward, 5′-CGGATCGGAGACGAGTTCA-3′; reverse, 5′-TTCAGCCTCGCGGTAATCA-3′); bid (forward, 5′- GAAGACGAGCTGCAGACAGATG-3′; reverse, 5′- TGGCTCTATTCTTCCTTGGTTGA-3′); bad (forward, 5′- GCAGGCACTGCAACACAGAT-3′; reverse, 5′- CTCCTTTGCCCAAGTTTCGAT-3′); bmf (forward, 5′- TACGCAACAACACCAGCAGAA-3′; reverse, 5′- CGAGGTTTTGAAGGAAGAGGAA-3′); dp5 (forward, 5′- TGGAGGAAGCTGGTTCCTGTT-3′; reverse, 5′- CAGCTCTTTACAATTCTGCTTCCTT-3′); cyclophilin (forward, 5′-CGCTTCCCAGATGAGAACTTCA-3′; reverse, 5- ACTGTGGTTATGAAGAACTGTGA-3′).

    Article Title: Decrease of the DNA methylation levels of the ADRB3 gene in leukocytes is related with serum folate in eutrophic adults
    Article Snippet: PCR was performed in a total volume of 20 μl containing: 1× buffer, 4 mM Mg2+ , 200 μM of each dNTPs (Qiagen), 250 nM of each primer, 5 μM SYTO® ), F:5′-TAGGTGATTTGGGAGATTTTTTTT-3′ and R:5′-CCCCTAACAACCCACTAATATTAAC-3′. .. The melting curves were normalized by calculation of the ‘line of best fit’ in between two normalization regions before and after the major fluorescence decrease representing the melting of the PCR product using the software provided with the HRM Software v2.0, provided by 7500 fast system.

    SYBR Green Assay:

    Article Title: Protein Phosphatase 2A Regulates bim Expression via the Akt/FKHRL1 Signaling Pathway in Amyloid-β Peptide-Induced Cerebrovascular Endothelial Cell Death
    Article Snippet: Equal amounts of total RNA (600 ng) were reverse transcribed with 1 μmol/L oligo (dT) and 0.5 mmol/L dNTPs (Qiagen), 10 U of Rnasin (Promega, Madison, WI), and 4 U of reverse transcriptase (Qiagen) for 1 h at 37°C. .. Ten nanograms of cDNA were subjected to PCR using the ABI Prism 7000 Sequence Detection System and SYBR Green PCR Master Mix (both from Applied Biosystems, Foster City, CA).

    RNA Extraction:

    Article Title: Aboveground Whitefly Infestation Modulates Transcriptional Levels of Anthocyanin Biosynthesis and Jasmonic Acid Signaling-Related Genes and Augments the Cope with Drought Stress of Maize
    Article Snippet: Paragraph title: Total RNA extraction and cDNA synthesis ... Synthesis of first-strand cDNA was performed from 1 μg of total RNA as a template with oligo-dT primer (Qiagen, CA, USA), dNTPs (Qiagen, CA, USA), and Moloney murine leukemia virus reverse transcriptase (MMLV-RT; Enzynomics, Daejeon, South Korea).

    Agarose Gel Electrophoresis:

    Article Title: A synonymous germline variant in a gene encoding a cell adhesion molecule is associated with cutaneous mast cell tumour development in Labrador and Golden Retrievers
    Article Snippet: A 1μl aliquot of a 1 : 100 dilution of each first round PCR product was used in a 20μl second round PCR reaction comprising 1 x HotStar Taq PCR buffer (Qiagen), 1 x Q-Solution (Qiagen), 0.5μM of both the ‘internal’ (B) forward and reverse primers , 0.3mM of each of 4 x dNTPs (Qiagen), 2 units of HotStar Taq DNA Polymerase (Qiagen), and 0.08 units of HotStar HiFidelity Taq DNA Polymerase (Qiagen). .. Second round PCR reaction products were analysed by 2% agarose gel electrophoresis (E14-15 Assay) and by 0.8% agarose gel electrophoresis (E15-17 Assay), respectively.

    Spectrophotometry:

    Article Title: Aboveground Whitefly Infestation Modulates Transcriptional Levels of Anthocyanin Biosynthesis and Jasmonic Acid Signaling-Related Genes and Augments the Cope with Drought Stress of Maize
    Article Snippet: RNA quality and quantity were confirmed by gel electrophoresis and a Nanodrop spectrophotometer (ND-1000, DE, USA) before use in subsequent analyses. .. Synthesis of first-strand cDNA was performed from 1 μg of total RNA as a template with oligo-dT primer (Qiagen, CA, USA), dNTPs (Qiagen, CA, USA), and Moloney murine leukemia virus reverse transcriptase (MMLV-RT; Enzynomics, Daejeon, South Korea).

    DNA Methylation Assay:

    Article Title: Decrease of the DNA methylation levels of the ADRB3 gene in leukocytes is related with serum folate in eutrophic adults
    Article Snippet: Paragraph title: Analysis of DNA methylation levels ... PCR was performed in a total volume of 20 μl containing: 1× buffer, 4 mM Mg2+ , 200 μM of each dNTPs (Qiagen), 250 nM of each primer, 5 μM SYTO® ), F:5′-TAGGTGATTTGGGAGATTTTTTTT-3′ and R:5′-CCCCTAACAACCCACTAATATTAAC-3′.

    Activation Assay:

    Article Title: Decrease of the DNA methylation levels of the ADRB3 gene in leukocytes is related with serum folate in eutrophic adults
    Article Snippet: PCR was performed in a total volume of 20 μl containing: 1× buffer, 4 mM Mg2+ , 200 μM of each dNTPs (Qiagen), 250 nM of each primer, 5 μM SYTO® ), F:5′-TAGGTGATTTGGGAGATTTTTTTT-3′ and R:5′-CCCCTAACAACCCACTAATATTAAC-3′. .. The PCR program consisted of an initial enzymatic activation at 95 °C for 10 min, followed by 50 cycles of 45 s at 95 °C, 45 s at 60 °C and 45 s at 72 °C and the final extension at 72 °C for 10 min.

    FACS:

    Article Title: Structural basis for enabling T-cell receptor diversity within biased virus-specific CD8+ T-cell responses
    Article Snippet: Single-cell sorting was performed using a BD FACS Aria. .. For transcription of cDNA from single, sorted cells, 5 μL of cDNA mix containing 0.25 μL of Sensiscript reverse transcriptase, 1× cDNA buffer, 0.5 mM dNTPs (Qiagen), 0.125 μg of oligo(dT) (Promega), 100 μg/mL gelatin (Roche), 100 μg/mL tRNA (Roche), 20 U RNase OUT (Invitrogen Life Technologies), and 0.1% Triton X-100 (Sigma-Aldrich) was added to wells and plates incubated at 37 °C for 90 min. Plates were stored at −80 °C and the TRAV21 and TRBV29 transcripts were later amplified by nested PCR and sequenced ( ).

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  • 96
    Qiagen dntps
    Determination of marker gene copy number by Southern blot hybridization . Southern-blot analysis of digested ( PstI ) genomic <t>DNA</t> from 9 randomly isolated paromomycin resistant mutants. A fragment of the AphVIII gene labeled with 32 <t>P-dNTPs</t> was used as probe. As shown, most transformants have a single copy of the integrated marker gene. Only the transformant represented in lane 4 may have two insertions (indicated by the two hybridizing bands). p.s., parental strain used to obtain the transformants.
    Dntps, supplied by Qiagen, used in various techniques. Bioz Stars score: 96/100, based on 867 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dntps/product/Qiagen
    Average 96 stars, based on 867 article reviews
    Price from $9.99 to $1999.99
    dntps - by Bioz Stars, 2020-02
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    79
    Qiagen residual dntps
    Confirmation of cleanup and enzymatic hydrolysis. ( A ) Removal of <t>dNTPs.</t> Samples were subjected to CE before and after a conventional <t>PCR</t> purification process. No peak was detected even in the 2-fold concentrated sample after purification. ( B ) Removal of primers. dNMP peaks resulting from residual primers were not detected in the 4-fold concentrated sample after purification. ( C ) CE profile of hydrolysis of a PCR product during a time course of phosphodiesterase treatment. dIMP was spiked as an internal standard for comparison. ( D ) Normalized dNMP peak intensities during a time course of phosphodiesterase treatment.
    Residual Dntps, supplied by Qiagen, used in various techniques. Bioz Stars score: 79/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/residual dntps/product/Qiagen
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    residual dntps - by Bioz Stars, 2020-02
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    Image Search Results


    Determination of marker gene copy number by Southern blot hybridization . Southern-blot analysis of digested ( PstI ) genomic DNA from 9 randomly isolated paromomycin resistant mutants. A fragment of the AphVIII gene labeled with 32 P-dNTPs was used as probe. As shown, most transformants have a single copy of the integrated marker gene. Only the transformant represented in lane 4 may have two insertions (indicated by the two hybridizing bands). p.s., parental strain used to obtain the transformants.

    Journal: Plant Methods

    Article Title: Reverse genetics in Chlamydomonas: a platform for isolating insertional mutants

    doi: 10.1186/1746-4811-7-24

    Figure Lengend Snippet: Determination of marker gene copy number by Southern blot hybridization . Southern-blot analysis of digested ( PstI ) genomic DNA from 9 randomly isolated paromomycin resistant mutants. A fragment of the AphVIII gene labeled with 32 P-dNTPs was used as probe. As shown, most transformants have a single copy of the integrated marker gene. Only the transformant represented in lane 4 may have two insertions (indicated by the two hybridizing bands). p.s., parental strain used to obtain the transformants.

    Article Snippet: PCR reactions using DNA from "superpools" as template were performed in a final volume of 25 μL and contained 0.4 pmoles of each primer, 0.2 mM of each of the dNTPs, 0.2 U of Taq DNA polymerase (Qiagen, Valencia, CA), 2.5 μL of 10 × Qiagen Taq DNA polymerase buffer, 5 μL Q solution (Qiagen, Valencia, CA), 100 ng of DNA template, and distilled water to make up the remainder of the 25 μL volume.

    Techniques: Marker, Southern Blot, Hybridization, Isolation, Labeling

    5FU incorporated into DNA is preferentially paired with adenine, followed by guanine. (A) Flowchart of procedure of in vitro analysis of binding partner of 5FU within DNA. (B-E) Frequencies of paired base with 5FU incorporated into DNA. One microgram of 5FdU:C containing dsDNA template PCR were utilized for both pH 8.2 or pH5.7 as a model of intracellular pH (pH i ) of tumor cells or normal cells in acidic tumor microenvironment with dNTP (B), only dCTP and dGTP (C), only dCTP, dGTP, and dATP (D), or only dCTP, dGTP, and dTTP (E). After TA cloning and total colony PCR, paired bases were analyzed by direct sequencing. The frequency of paired bases was calculated as the number of each base where 5FdU was inserted by total number of colonies ( N ≥ 24) with an informative sequence at the 5FdU site. 5FU within DNA is predominantly paired with adenine when both dGTP and dATP are available (B, D), whereas guanine is preferentially paired with 5FU when dATP is absent (C, E).

    Journal: Mutation research

    Article Title: Acidic tumor microenvironment downregulates hMLH1 but does not diminish 5-fluorouracil chemosensitivity

    doi: 10.1016/j.mrfmmm.2013.04.006

    Figure Lengend Snippet: 5FU incorporated into DNA is preferentially paired with adenine, followed by guanine. (A) Flowchart of procedure of in vitro analysis of binding partner of 5FU within DNA. (B-E) Frequencies of paired base with 5FU incorporated into DNA. One microgram of 5FdU:C containing dsDNA template PCR were utilized for both pH 8.2 or pH5.7 as a model of intracellular pH (pH i ) of tumor cells or normal cells in acidic tumor microenvironment with dNTP (B), only dCTP and dGTP (C), only dCTP, dGTP, and dATP (D), or only dCTP, dGTP, and dTTP (E). After TA cloning and total colony PCR, paired bases were analyzed by direct sequencing. The frequency of paired bases was calculated as the number of each base where 5FdU was inserted by total number of colonies ( N ≥ 24) with an informative sequence at the 5FdU site. 5FU within DNA is predominantly paired with adenine when both dGTP and dATP are available (B, D), whereas guanine is preferentially paired with 5FU when dATP is absent (C, E).

    Article Snippet: Purified genomic DNA was used for a PCR reaction mixture with 10 mM Tris–HCl [pH 8.4], 50 mM KCl, 1.5 mM MgCl2 , 200 μM dNTP, 5U of HotStarTaq DNA polymerase (QIAGEN), 0.25 μM forward primer (5′-AGAACCCACTGCTTACTGGC-3′) and reverse primer (5′-ATGGCTGGCAACTAGAAGGC-3/ ) whose sequence is identical with each end of 5FdU:G, T:G, or C:G matched dsDNA.

    Techniques: In Vitro, Binding Assay, Polymerase Chain Reaction, TA Cloning, Sequencing

    Confirmation of cleanup and enzymatic hydrolysis. ( A ) Removal of dNTPs. Samples were subjected to CE before and after a conventional PCR purification process. No peak was detected even in the 2-fold concentrated sample after purification. ( B ) Removal of primers. dNMP peaks resulting from residual primers were not detected in the 4-fold concentrated sample after purification. ( C ) CE profile of hydrolysis of a PCR product during a time course of phosphodiesterase treatment. dIMP was spiked as an internal standard for comparison. ( D ) Normalized dNMP peak intensities during a time course of phosphodiesterase treatment.

    Journal: Nucleic Acids Research

    Article Title: Rapid quantification of DNA methylation through dNMP analysis following bisulfite-PCR

    doi: 10.1093/nar/gkl257

    Figure Lengend Snippet: Confirmation of cleanup and enzymatic hydrolysis. ( A ) Removal of dNTPs. Samples were subjected to CE before and after a conventional PCR purification process. No peak was detected even in the 2-fold concentrated sample after purification. ( B ) Removal of primers. dNMP peaks resulting from residual primers were not detected in the 4-fold concentrated sample after purification. ( C ) CE profile of hydrolysis of a PCR product during a time course of phosphodiesterase treatment. dIMP was spiked as an internal standard for comparison. ( D ) Normalized dNMP peak intensities during a time course of phosphodiesterase treatment.

    Article Snippet: PCR products were cleaned up to remove residual dNTPs and primers using a conventional PCR purification kit (Qiagen, Hilden, Germany).

    Techniques: Polymerase Chain Reaction, Purification