Reverse Transcription:Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR 2.5.3. .. Quantitative real-time PCR The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR .. The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Incubation:Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR 2.5.3. .. Quantitative real-time PCR The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR .. The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Marker:Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR 2.5.3. .. Quantitative real-time PCR The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR .. The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Expressing:Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR 2.5.3. .. Quantitative real-time PCR The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR .. The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Quantitative RT-PCR:Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR 2.5.3. .. Quantitative real-time PCR The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR .. The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Sequencing:Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR 2.5.3. .. Quantitative real-time PCR The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR .. The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Concentration Assay:Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR 2.5.3. .. Quantitative real-time PCR The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR .. The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Real-time Polymerase Chain Reaction:Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR 2.5.3. .. Quantitative real-time PCR The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
Article Title: Mesenchymal stem cell function on hybrid organic/inorganic microparticles in vitro
Article Snippet: .. The mastermix (15 μl), containing nuclease-free water, MMLV 5 X buffer (Promega, Cat. No. M531A), 10 mM dNTP and reverse transcriptase (Promega, Cat. No. M314A) (9 : 4:1 : 1 ratio) was added to each tube, and the tubes were incubated at room temperature for 5 min and then placed at 42 °C for 1 h, followed by 70 °C for 15 min. OB-specific gene marker expression, such as alkaline phosphatase ( ALP ), collagen I ( COLLI ) and osteocalcin ( OCN ), was assessed by real-time RT–PCR, using primers ( Fu et al ., 2007 ; Moerman et al ., 2004 ) as described in . table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Gene name Primer sequence Number of base pairs Primer concentration (nM) ALP Forward 5′-GTGCCAGAGAAAGAGAGAGAC-3′ 21 32.6 Reverse 5′-GACGCCCATACCATCTCC-3′ 18 37.1 OCN Forward 5′-GAGTCTGACAAAGCCTTCA-3′ 19 35.4 Reverse 5′-AGCCATACTGGTCTGATAG-3′ 19 36.4 COLLI Forward 5′-ACTGTCCCAACCCCCAAAG-3′ 19 33 Reverse 5′-CGTATTCTTCCGGGCAGAAA-3′ 20 33.3 GAPDH Forward 5′-GTCGGTGTGAACGGATTTG-3′ 19 33.1 Reverse 5′-GAACATGTAGACCATGTAGTTG-3′ 22 34.4 Open in a separate window Primer sequences used for quantitative real-time PCR .. The DNA samples were amplified using lightcycler Quantitative real-time PCR (Roche Diagnostics).
|
