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  • 99
    Qiagen rnase free water
    Evaluation of the miRNeasy (Qiagen) and miRCURY (Exiqon) extraction kits. ( A ) Eight healthy blood donors were used. <t>RNA</t> was extracted from 200 µL serum using the miRNeasy RNA isolation kit (Qiagen) and miRCURY RNA isolation kit Biofuids (Exiqon) following the manufacturer’s instructions, and eluted in 14 µL and 50 µL of <t>RNase</t> free water respectively. TaqMan microRNA quantitative reverse transcription polymerase chain reaction (qRT-PCR) (Applied Biosystems, Waltham, MA, USA) was carried out against miR-423-3p, a normalizer recommended by Exiqon. Cq values for miR-423-3p obtained from RNA extracted using the miRNeasy RNA isolation kit (Red) verses Cq values obtained from RNA extracted using the miRCURY RNA isolation kit Biofluids (Green) are displayed. ( B ). *** p
    Rnase Free Water, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 3400 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rnase free water/product/Qiagen
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    rnase free water - by Bioz Stars, 2020-04
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    99
    Millipore rnase dnase free water
    Evaluation of the miRNeasy (Qiagen) and miRCURY (Exiqon) extraction kits. ( A ) Eight healthy blood donors were used. <t>RNA</t> was extracted from 200 µL serum using the miRNeasy RNA isolation kit (Qiagen) and miRCURY RNA isolation kit Biofuids (Exiqon) following the manufacturer’s instructions, and eluted in 14 µL and 50 µL of <t>RNase</t> free water respectively. TaqMan microRNA quantitative reverse transcription polymerase chain reaction (qRT-PCR) (Applied Biosystems, Waltham, MA, USA) was carried out against miR-423-3p, a normalizer recommended by Exiqon. Cq values for miR-423-3p obtained from RNA extracted using the miRNeasy RNA isolation kit (Red) verses Cq values obtained from RNA extracted using the miRCURY RNA isolation kit Biofluids (Green) are displayed. ( B ). *** p
    Rnase Dnase Free Water, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 150 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa rnase free water
    Overview of RT-RamDA and single-cell RamDA-seq. a Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a RNA template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of <t>RNase</t> H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the <t>T4</t> gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I. b Relative yield of cDNA molecules using RT-qPCR ( n = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog , Pou5f1 , Zfp42 , and Sox2 ) and three housekeeping ( Gnb2l1 , Atp5a1 , and Tubb5 ) genes using a conventional method (−) as a standard. c Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section. d Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in b and d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile. e Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in d and e . Transcripts were annotated by GENCODE gene annotation (vM9)
    Rnase Free Water, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 731 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Millipore rnase free water
    Overview of RT-RamDA and single-cell RamDA-seq. a Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a RNA template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of <t>RNase</t> H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the <t>T4</t> gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I. b Relative yield of cDNA molecules using RT-qPCR ( n = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog , Pou5f1 , Zfp42 , and Sox2 ) and three housekeeping ( Gnb2l1 , Atp5a1 , and Tubb5 ) genes using a conventional method (−) as a standard. c Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section. d Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in b and d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile. e Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in d and e . Transcripts were annotated by GENCODE gene annotation (vM9)
    Rnase Free Water, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 1154 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Promega rnase free water
    Overview of RT-RamDA and single-cell RamDA-seq. a Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a RNA template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of <t>RNase</t> H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the <t>T4</t> gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I. b Relative yield of cDNA molecules using RT-qPCR ( n = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog , Pou5f1 , Zfp42 , and Sox2 ) and three housekeeping ( Gnb2l1 , Atp5a1 , and Tubb5 ) genes using a conventional method (−) as a standard. c Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section. d Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in b and d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile. e Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in d and e . Transcripts were annotated by GENCODE gene annotation (vM9)
    Rnase Free Water, supplied by Promega, used in various techniques. Bioz Stars score: 97/100, based on 1012 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Mediatech rnase free water
    Overview of RT-RamDA and single-cell RamDA-seq. a Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a RNA template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of <t>RNase</t> H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the <t>T4</t> gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I. b Relative yield of cDNA molecules using RT-qPCR ( n = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog , Pou5f1 , Zfp42 , and Sox2 ) and three housekeeping ( Gnb2l1 , Atp5a1 , and Tubb5 ) genes using a conventional method (−) as a standard. c Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section. d Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in b and d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile. e Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in d and e . Transcripts were annotated by GENCODE gene annotation (vM9)
    Rnase Free Water, supplied by Mediatech, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher rnase free water
    Overview of RT-RamDA and single-cell RamDA-seq. a Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a <t>RNA</t> template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of <t>RNase</t> H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the T4 gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I. b Relative yield of cDNA molecules using RT-qPCR ( n = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog , Pou5f1 , Zfp42 , and Sox2 ) and three housekeeping ( Gnb2l1 , Atp5a1 , and Tubb5 ) genes using a conventional method (−) as a standard. c Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section. d Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in b and d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile. e Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in d and e . Transcripts were annotated by GENCODE gene annotation (vM9)
    Rnase Free Water, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 9992 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Beijing CWBio rnase free water
    Overview of RT-RamDA and single-cell RamDA-seq. a Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a <t>RNA</t> template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of <t>RNase</t> H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the T4 gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I. b Relative yield of cDNA molecules using RT-qPCR ( n = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog , Pou5f1 , Zfp42 , and Sox2 ) and three housekeeping ( Gnb2l1 , Atp5a1 , and Tubb5 ) genes using a conventional method (−) as a standard. c Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section. d Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in b and d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile. e Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in d and e . Transcripts were annotated by GENCODE gene annotation (vM9)
    Rnase Free Water, supplied by Beijing CWBio, used in various techniques. Bioz Stars score: 91/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Carl Roth GmbH rnase free water
    Overview of RT-RamDA and single-cell RamDA-seq. a Schematic diagram of RT-RamDA. 1. RT primers (oligo-dT and not-so-random primers) anneal to a <t>RNA</t> template. 2. Complementary DNA (cDNA) is synthesized by the RNA-dependent DNA polymerase activity of <t>RNase</t> H minus reverse transcriptase (RTase). 3. Endonuclease (DNase I) selectively nicks the cDNA of the RNA:cDNA hybrid strand. 4. The 3′ cDNA strand is displaced by the strand displacement activity of RTase mediated by the T4 gene 32 protein (gp32), starting from the nick randomly introduced by DNase I. cDNA is amplified as a displaced strand and protected by gp32 from DNase I. b Relative yield of cDNA molecules using RT-qPCR ( n = 4). Mouse ESC total RNA (10 pg) was used as a template, and 1/10 the amount of cDNA was used for qPCR. The relative yield was calculated by averaging the amplification efficiency of four mESC ( Nanog , Pou5f1 , Zfp42 , and Sox2 ) and three housekeeping ( Gnb2l1 , Atp5a1 , and Tubb5 ) genes using a conventional method (−) as a standard. c Schematic diagram of RamDA-seq and C1-RamDA-seq. For details, please refer to the Methods section. d Number of detected transcripts with twofold or lower expression changes against rdRNA-seq (count ≥ 10). For the boxplots in b and d , the center line, and lower and upper bounds of each box represent the median, and first and third quartiles, respectively. The lower (upper) whisker extends to smallest (largest) values no further than 1.5 × interquartile range (IQR) from the first (third) quartile. e Squared coefficient of variation of the read count. All conditions were adjusted, and 10 million reads were used in d and e . Transcripts were annotated by GENCODE gene annotation (vM9)
    Rnase Free Water, supplied by Carl Roth GmbH, used in various techniques. Bioz Stars score: 93/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Roche rnase free water
    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
    Rnase Free Water, supplied by Roche, used in various techniques. Bioz Stars score: 97/100, based on 683 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Bio Basic Canada rnase free water
    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
    Rnase Free Water, supplied by Bio Basic Canada, used in various techniques. Bioz Stars score: 96/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TransGen biotech co rnase free water
    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
    Rnase Free Water, supplied by TransGen biotech co, used in various techniques. Bioz Stars score: 95/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Fisher Scientific rnase free water
    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
    Rnase Free Water, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 96/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    GE Healthcare rnase free water
    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
    Rnase Free Water, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 96/100, based on 273 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    RPI Inc rnase free water
    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
    Rnase Free Water, supplied by RPI Inc, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Avantor rnase free water
    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
    Rnase Free Water, supplied by Avantor, used in various techniques. Bioz Stars score: 92/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Wisent Corporation rnase free water
    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
    Rnase Free Water, supplied by Wisent Corporation, used in various techniques. Bioz Stars score: 91/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Tel Test Inc rnase free water
    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) <t>RNase</t> R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated <t>cDNA</t> employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.
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    Image Search Results


    Evaluation of the miRNeasy (Qiagen) and miRCURY (Exiqon) extraction kits. ( A ) Eight healthy blood donors were used. RNA was extracted from 200 µL serum using the miRNeasy RNA isolation kit (Qiagen) and miRCURY RNA isolation kit Biofuids (Exiqon) following the manufacturer’s instructions, and eluted in 14 µL and 50 µL of RNase free water respectively. TaqMan microRNA quantitative reverse transcription polymerase chain reaction (qRT-PCR) (Applied Biosystems, Waltham, MA, USA) was carried out against miR-423-3p, a normalizer recommended by Exiqon. Cq values for miR-423-3p obtained from RNA extracted using the miRNeasy RNA isolation kit (Red) verses Cq values obtained from RNA extracted using the miRCURY RNA isolation kit Biofluids (Green) are displayed. ( B ). *** p

    Journal: Biomolecules

    Article Title: Circulating MicroRNA Biomarkers in Melanoma: Tools and Challenges in Personalised Medicine

    doi: 10.3390/biom8020021

    Figure Lengend Snippet: Evaluation of the miRNeasy (Qiagen) and miRCURY (Exiqon) extraction kits. ( A ) Eight healthy blood donors were used. RNA was extracted from 200 µL serum using the miRNeasy RNA isolation kit (Qiagen) and miRCURY RNA isolation kit Biofuids (Exiqon) following the manufacturer’s instructions, and eluted in 14 µL and 50 µL of RNase free water respectively. TaqMan microRNA quantitative reverse transcription polymerase chain reaction (qRT-PCR) (Applied Biosystems, Waltham, MA, USA) was carried out against miR-423-3p, a normalizer recommended by Exiqon. Cq values for miR-423-3p obtained from RNA extracted using the miRNeasy RNA isolation kit (Red) verses Cq values obtained from RNA extracted using the miRCURY RNA isolation kit Biofluids (Green) are displayed. ( B ). *** p

    Article Snippet: The difference in Cq observed between kits in a is expected as RNA is eluted in 3.57× the amount of RNase free water using the miRCURY RNA isolation kit (Exiqon) compared to the miRNeasy RNA isolation kit (Qiagen), resulting in a lower concentration.

    Techniques: Isolation, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

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

    Journal: Nature Communications

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

    doi: 10.1038/s41467-018-02866-0

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

    Article Snippet: One microliter of RT-RamDA mix (2.5× PrimeScript Buffer, 0.6 pmol oligo(dT)18, 8 pmol 1st-NSRs, 100 ng of T4 gene 32 protein, and 3× PrimeScript enzyme mix in RNase-free water) was added to 2 µL of the digested lysates.

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

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

    Journal: Nature Communications

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

    doi: 10.1038/s41467-018-02866-0

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

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

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

    Mutational analysis of the predicted stem-loop structure in the N-SL2 . (A) Strategic representation of RT-PCR used to detect (-) gRNA, (+) sg mRNA7 and (-) sg mRNA7. The positions are according to APRRSV stain (GenBank: GQ330474 ) and all primer sequences are listed in Table 1. pAS was a non-replicative control which was absence of gene ORF1a and ORF1b (1688-13118) in full-length cDNA clone. (B) Schematic representation of the mutations introduced into the N-SL2 structure. The loop was enlarged as described in Figure 1, and mutants L-LL and L-RR were generated by overlapping PCR mutagenesis. L-RL was generated by combining the right and left arm sequences of the L-LL and L-RR, respectively, such that the overall structure of N-SL2 was restored. All the mutated nucleotides (lowercase) are highlighted in gray shading. The stem mutants, S-LL and S-RR, were generated by overlapping PCR such that one arm sequence was replaced with that of the opposite arm. The double mutant, S-RL, was generated by combining the mutations in the left and right arms such that the overall structure was restored. All mutant sequences are shown as lowercase. (C) RT-PCR of RNAs extracted from pAS and WT transfected cells at 24 hours after transfection. DNase I and RNase A were used to omit template DNA and the reverse transcriptase. The primers were nested RT-PCR primers as same as (-) gRNA detection. A 2-kbp ladder was used as a molecular size marker. The numbers indicated the lane No. (D) RT-PCR analysis of the mutants. Total cellular RNAs were extracted from mutant plasmids-transfected from BHK-21 cells at 24 hours post-transfection. β-actin is a marker for the level of intracellular RNA isolation, and pAS is a non-replicative control.

    Journal: Virology Journal

    Article Title: A 5'-proximal Stem-loop Structure of 5' Untranslated Region of Porcine Reproductive and Respiratory Syndrome Virus Genome Is Key for Virus Replication

    doi: 10.1186/1743-422X-8-172

    Figure Lengend Snippet: Mutational analysis of the predicted stem-loop structure in the N-SL2 . (A) Strategic representation of RT-PCR used to detect (-) gRNA, (+) sg mRNA7 and (-) sg mRNA7. The positions are according to APRRSV stain (GenBank: GQ330474 ) and all primer sequences are listed in Table 1. pAS was a non-replicative control which was absence of gene ORF1a and ORF1b (1688-13118) in full-length cDNA clone. (B) Schematic representation of the mutations introduced into the N-SL2 structure. The loop was enlarged as described in Figure 1, and mutants L-LL and L-RR were generated by overlapping PCR mutagenesis. L-RL was generated by combining the right and left arm sequences of the L-LL and L-RR, respectively, such that the overall structure of N-SL2 was restored. All the mutated nucleotides (lowercase) are highlighted in gray shading. The stem mutants, S-LL and S-RR, were generated by overlapping PCR such that one arm sequence was replaced with that of the opposite arm. The double mutant, S-RL, was generated by combining the mutations in the left and right arms such that the overall structure was restored. All mutant sequences are shown as lowercase. (C) RT-PCR of RNAs extracted from pAS and WT transfected cells at 24 hours after transfection. DNase I and RNase A were used to omit template DNA and the reverse transcriptase. The primers were nested RT-PCR primers as same as (-) gRNA detection. A 2-kbp ladder was used as a molecular size marker. The numbers indicated the lane No. (D) RT-PCR analysis of the mutants. Total cellular RNAs were extracted from mutant plasmids-transfected from BHK-21 cells at 24 hours post-transfection. β-actin is a marker for the level of intracellular RNA isolation, and pAS is a non-replicative control.

    Article Snippet: RNAs were suspended in DNase/RNase-free water, and quantified by NanoDrop® ND-1000 (Thermo Fisher Scientific Inc.).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Staining, Generated, Polymerase Chain Reaction, Mutagenesis, Sequencing, Transfection, Marker, Isolation

    Particle size analysis of the TOP complex. T-oligo and PVBLG solutions were prepared at a DNA to polymer weight ratio of 15:1 in DNAse/RNAse-free distilled water. T-oligo and PVBLG solutions were then mixed and incubated at room temperature for 20 minutes. The particle size was analyzed by dynamic light scattering. The results obtained are shown in NICOMP (dynamic light scattering) distribution, indicating that the TOP complex had a mean diameter of 147.3 nm. PVBLG or T-oligo alone was used as the control. Abbreviation: TOP complex, T-oligo-PVBLG nanocomplex.

    Journal: International Journal of Nanomedicine

    Article Title: Novel delivery system for T-oligo using a nanocomplex formed with an alpha helical peptide for melanoma therapy

    doi: 10.2147/IJN.S55133

    Figure Lengend Snippet: Particle size analysis of the TOP complex. T-oligo and PVBLG solutions were prepared at a DNA to polymer weight ratio of 15:1 in DNAse/RNAse-free distilled water. T-oligo and PVBLG solutions were then mixed and incubated at room temperature for 20 minutes. The particle size was analyzed by dynamic light scattering. The results obtained are shown in NICOMP (dynamic light scattering) distribution, indicating that the TOP complex had a mean diameter of 147.3 nm. PVBLG or T-oligo alone was used as the control. Abbreviation: TOP complex, T-oligo-PVBLG nanocomplex.

    Article Snippet: Particle size and zeta potential analysis T-oligo and PVBLG solutions were prepared separately in DNAse/RNAse-free distilled water (Invitrogen, Grand Island, NY, USA) and complexed at various weight ratios.

    Techniques: Particle Size Analysis, Incubation

    Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) RNase R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated cDNA employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.

    Journal: Nucleic Acids Research

    Article Title: Human Survival Motor Neuron genes generate a vast repertoire of circular RNAs

    doi: 10.1093/nar/gkz034

    Figure Lengend Snippet: Linear SMN transcripts showing inclusion of novel exons 9, 10, 11 and 12. ( A ) An overview of the 3′ portion of the SMN pre-mRNA. Splicing events are indicated by dotted lines. Exons are depicted as colored shapes, and introns are shown as lines/broken lines. Exon sizes are indicated by numbers in black below exons and intron sizes are indicated by numbers in gray above introns. Names, assigned numbers and the locations of primers used to amplify linear SMN transcripts are presented. ‘Δ’ indicates SMN transcripts with skipped exon(s). ( B, C ) Detection of novel exons 9, 10, 11 and 12 without (−) and with (+) RNase R treatment. Primers used are shown at the top of each gel. After purification of total RNA, we isolated intact poly(A) + RNA using magnetic beads and generated cDNA employing oligo(dT) 12–18 primer. The identities of bands marked on the left of each gel were determined by cloning and sequencing. Abbreviations: NS, non-specific.

    Article Snippet: The cDNA was further diluted 20-fold with RNase-free water, and 3 μl was then used in a 20 μl of PCR reaction using FastStart Universal SYBR Green Master mix (ROX) (Roche). qPCR was performed using a Stratagene Mx3005 qPCR system (Agilent Technologies).

    Techniques: Purification, Isolation, Magnetic Beads, Generated, Clone Assay, Sequencing