mirvana mirna isolation kit  (Thermo Fisher)


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    Name:
    mirVana miRNA Isolation Kit
    Description:
    The mirVana miRNA Isolation Kit uses a rapid procedure to isolate small RNAs from tissue and cells using an efficient glass fiber filter (GFF)-based method. The method isolates total RNA ranging in size from kilobases down to 10-mers. Each kit contains sufficient reagents and consumables for either 40 isolations of total RNA (including small RNAs), or 20 separate large and small RNA fractions.Features of this kit include:• Efficient isolation of small RNA-containing total RNA• Enrich for small RNA (<200 nt) to increase sensitivity in downstream analyses• Simple 30 minute procedure• Ideal for miRNA, siRNA, shRNA, and snRNA analysis• Compatible with virtually all cell and tissue typesEfficient recovery of small RNACurrent RNA purification procedures rely on organic extraction, followed by alcohol precipitation or adsorption of nucleic acid molecules on a GFF or silicate matrix. Unfortunately, the former procedure requires alcohol precipitation, which is time consuming and inefficiently recovers small RNAs. The latter procedure typically results in the loss of substantial amounts of small RNA. The mirVana miRNA Isolation Kit uses organic extraction followed by purification on a GFF under specialized binding and wash conditions. As a result, the kit effectively recovers all RNA—from large mRNA and ribosomal RNA down to 10-mers—from virtually all cell and tissue types. The mirVana miRNA Isolation Kit enables the isolation of small RNA-containing total RNA from samples consisting of 102–107 cultured cells or 0.5–250 mg tissue.Enrich for small RNAAlthough the mirVana miRNA Isolation Kit efficiently purifies all RNA larger than 10 nt, it also includes procedures for isolating RNA fractions, specifically enriched or depleted in small RNA species. Through the enrichment procedure included in the mirVana miRNA Isolation Kit, RNA molecules of ~200 nt and less can be efficiently purified away from the larger RNA species. The resulting RNA preparation is highly enriched for miRNAs, siRNAs, and/or snRNAs. Small RNA enrichment using the mirVana miRNA Isolation Kit procedure allows more sensitive small RNA detection with less background as compared to the same assay used with total RNA.Accessory productThe mirVana miRNA Isolation Kit without Phenol is available for customers concerned about the cost of shipping organic chemicals such as Acid Phenol:Chloroform, which is included in the standard kit.
    Catalog Number:
    AM1560
    Price:
    None
    Applications:
    DNA & RNA Purification & Analysis|RNAi|RNAi, Epigenetics & Non-Coding RNA Research|RNA Extraction|miRNA Isolation|miRNA & Non-Coding RNA Analysis|microRNA Isolation|siRNA
    Size:
    40 preps
    Category:
    Kits and Assays, DNA⁄RNA Purification Kits & Reagents, miRNA Isolation Kits
    Score:
    85
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    Structured Review

    Thermo Fisher mirvana mirna isolation kit
    Schematic of HSV-1 and organization of the LAT locus. (A) The LAT locus is present in the unique long (UL) and unique short (US) regions. The primary 8.3-kb LAT is shown as a long arrow. The stable 2-kb LAT is shown as a solid rectangle. The LAT TATA box is denoted by TATA, and the small arrow and +1 indicate the start of LAT transcription (genomic nt 118801). The LAT promoter (LAP) is denoted by the gray rectangle. The relative locations of mRNAs encoding ICP0 and ICP34.5 are shown for reference. Expression of the first 1.5 kb of LAT coding sequences (+1,500) is sufficient for reactivation from latency . The relative locations of the two LAT sRNAs that were previously identified are denoted by asterisks . (B) Nucleotide sequences of the LAT sRNAs . The underlined ATGs correspond to the initiating ATGs of ORF4 (LAT sRNA1) or ORF8 (LAT sRNA2) . (C) Female Swiss Webster mice were infected with dLAT2903R (lane R) or dLAT2903 (lane 2903). At 30 days after infection, TG were extracted and sRNA was prepared by using the <t>miRNA</t> isolation kit (miVana; Ambion) according to the manufacturer's instructions. RT-PCR using adaptor primers and LAT-specific primers was used to examine sRNA expression, as described in Materials and Methods. (D) Double-stranded DNA corresponding to the respective sRNAs was synthesized and cloned into the siRNA expression plasmid, pSilencer 2.1-U6 neo (Ambion). neuro-2a cells were transfected with 5 μg of a plasmid expressing LAT sRNA1 (lane 1), LAT sRNA2 (lane 2), or the empty pSilencer 2.1-U6 neo (lane E). At 40 h after transfection, sRNA was prepared using the <t>mirVana</t> miRNA isolation kit (Ambion) according to the manufacturer's instructions, and the expression of LAT sRNA was examined by RT-PCR. Following purification of sRNA by the mirVana kit, total sRNA (1 μg) was electrophoresed on a 2% agarose gel to confirm that similar levels of RNA were used for RT-PCR amplification.
    The mirVana miRNA Isolation Kit uses a rapid procedure to isolate small RNAs from tissue and cells using an efficient glass fiber filter (GFF)-based method. The method isolates total RNA ranging in size from kilobases down to 10-mers. Each kit contains sufficient reagents and consumables for either 40 isolations of total RNA (including small RNAs), or 20 separate large and small RNA fractions.Features of this kit include:• Efficient isolation of small RNA-containing total RNA• Enrich for small RNA (<200 nt) to increase sensitivity in downstream analyses• Simple 30 minute procedure• Ideal for miRNA, siRNA, shRNA, and snRNA analysis• Compatible with virtually all cell and tissue typesEfficient recovery of small RNACurrent RNA purification procedures rely on organic extraction, followed by alcohol precipitation or adsorption of nucleic acid molecules on a GFF or silicate matrix. Unfortunately, the former procedure requires alcohol precipitation, which is time consuming and inefficiently recovers small RNAs. The latter procedure typically results in the loss of substantial amounts of small RNA. The mirVana miRNA Isolation Kit uses organic extraction followed by purification on a GFF under specialized binding and wash conditions. As a result, the kit effectively recovers all RNA—from large mRNA and ribosomal RNA down to 10-mers—from virtually all cell and tissue types. The mirVana miRNA Isolation Kit enables the isolation of small RNA-containing total RNA from samples consisting of 102–107 cultured cells or 0.5–250 mg tissue.Enrich for small RNAAlthough the mirVana miRNA Isolation Kit efficiently purifies all RNA larger than 10 nt, it also includes procedures for isolating RNA fractions, specifically enriched or depleted in small RNA species. Through the enrichment procedure included in the mirVana miRNA Isolation Kit, RNA molecules of ~200 nt and less can be efficiently purified away from the larger RNA species. The resulting RNA preparation is highly enriched for miRNAs, siRNAs, and/or snRNAs. Small RNA enrichment using the mirVana miRNA Isolation Kit procedure allows more sensitive small RNA detection with less background as compared to the same assay used with total RNA.Accessory productThe mirVana miRNA Isolation Kit without Phenol is available for customers concerned about the cost of shipping organic chemicals such as Acid Phenol:Chloroform, which is included in the standard kit.
    https://www.bioz.com/result/mirvana mirna isolation kit/product/Thermo Fisher
    Average 99 stars, based on 2899 article reviews
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    Images

    1) Product Images from "Two Small RNAs Encoded within the First 1.5 Kilobases of the Herpes Simplex Virus Type 1 Latency-Associated Transcript Can Inhibit Productive Infection and Cooperate To Inhibit Apoptosis "

    Article Title: Two Small RNAs Encoded within the First 1.5 Kilobases of the Herpes Simplex Virus Type 1 Latency-Associated Transcript Can Inhibit Productive Infection and Cooperate To Inhibit Apoptosis

    Journal: Journal of Virology

    doi: 10.1128/JVI.00871-09

    Schematic of HSV-1 and organization of the LAT locus. (A) The LAT locus is present in the unique long (UL) and unique short (US) regions. The primary 8.3-kb LAT is shown as a long arrow. The stable 2-kb LAT is shown as a solid rectangle. The LAT TATA box is denoted by TATA, and the small arrow and +1 indicate the start of LAT transcription (genomic nt 118801). The LAT promoter (LAP) is denoted by the gray rectangle. The relative locations of mRNAs encoding ICP0 and ICP34.5 are shown for reference. Expression of the first 1.5 kb of LAT coding sequences (+1,500) is sufficient for reactivation from latency . The relative locations of the two LAT sRNAs that were previously identified are denoted by asterisks . (B) Nucleotide sequences of the LAT sRNAs . The underlined ATGs correspond to the initiating ATGs of ORF4 (LAT sRNA1) or ORF8 (LAT sRNA2) . (C) Female Swiss Webster mice were infected with dLAT2903R (lane R) or dLAT2903 (lane 2903). At 30 days after infection, TG were extracted and sRNA was prepared by using the miRNA isolation kit (miVana; Ambion) according to the manufacturer's instructions. RT-PCR using adaptor primers and LAT-specific primers was used to examine sRNA expression, as described in Materials and Methods. (D) Double-stranded DNA corresponding to the respective sRNAs was synthesized and cloned into the siRNA expression plasmid, pSilencer 2.1-U6 neo (Ambion). neuro-2a cells were transfected with 5 μg of a plasmid expressing LAT sRNA1 (lane 1), LAT sRNA2 (lane 2), or the empty pSilencer 2.1-U6 neo (lane E). At 40 h after transfection, sRNA was prepared using the mirVana miRNA isolation kit (Ambion) according to the manufacturer's instructions, and the expression of LAT sRNA was examined by RT-PCR. Following purification of sRNA by the mirVana kit, total sRNA (1 μg) was electrophoresed on a 2% agarose gel to confirm that similar levels of RNA were used for RT-PCR amplification.
    Figure Legend Snippet: Schematic of HSV-1 and organization of the LAT locus. (A) The LAT locus is present in the unique long (UL) and unique short (US) regions. The primary 8.3-kb LAT is shown as a long arrow. The stable 2-kb LAT is shown as a solid rectangle. The LAT TATA box is denoted by TATA, and the small arrow and +1 indicate the start of LAT transcription (genomic nt 118801). The LAT promoter (LAP) is denoted by the gray rectangle. The relative locations of mRNAs encoding ICP0 and ICP34.5 are shown for reference. Expression of the first 1.5 kb of LAT coding sequences (+1,500) is sufficient for reactivation from latency . The relative locations of the two LAT sRNAs that were previously identified are denoted by asterisks . (B) Nucleotide sequences of the LAT sRNAs . The underlined ATGs correspond to the initiating ATGs of ORF4 (LAT sRNA1) or ORF8 (LAT sRNA2) . (C) Female Swiss Webster mice were infected with dLAT2903R (lane R) or dLAT2903 (lane 2903). At 30 days after infection, TG were extracted and sRNA was prepared by using the miRNA isolation kit (miVana; Ambion) according to the manufacturer's instructions. RT-PCR using adaptor primers and LAT-specific primers was used to examine sRNA expression, as described in Materials and Methods. (D) Double-stranded DNA corresponding to the respective sRNAs was synthesized and cloned into the siRNA expression plasmid, pSilencer 2.1-U6 neo (Ambion). neuro-2a cells were transfected with 5 μg of a plasmid expressing LAT sRNA1 (lane 1), LAT sRNA2 (lane 2), or the empty pSilencer 2.1-U6 neo (lane E). At 40 h after transfection, sRNA was prepared using the mirVana miRNA isolation kit (Ambion) according to the manufacturer's instructions, and the expression of LAT sRNA was examined by RT-PCR. Following purification of sRNA by the mirVana kit, total sRNA (1 μg) was electrophoresed on a 2% agarose gel to confirm that similar levels of RNA were used for RT-PCR amplification.

    Techniques Used: Expressing, Mouse Assay, Infection, Isolation, Reverse Transcription Polymerase Chain Reaction, Synthesized, Clone Assay, Plasmid Preparation, Transfection, Purification, Agarose Gel Electrophoresis, Amplification

    2) Product Images from "The Bioactivity of D-/L-Isonucleoside- and 2′-Deoxyinosine-Incorporated Aptamer AS1411s Including DNA Replication/MicroRNA Expression"

    Article Title: The Bioactivity of D-/L-Isonucleoside- and 2′-Deoxyinosine-Incorporated Aptamer AS1411s Including DNA Replication/MicroRNA Expression

    Journal: Molecular Therapy. Nucleic Acids

    doi: 10.1016/j.omtn.2017.09.010

    Modulation of MicroRNA Expression by FCL-II and Suppression Effect of FCL-I/II on MCF-7 Xenografts (A) For TaqMan PCR, the total RNA was isolated using the mirVana miRNA Isolation Kit (Life Technologies Corporation, USA). RT-PCR was performed using the TaqMan MicroRNA Assay (Life Technologies Corporation, USA) and the Mx3005P QPCR System (Agilent Technologies, Inc., CA, USA). U6 gene quantification was used as the control. The results are shown as a mean of three separate experiments with SD. (B) Bilaterally ovariectomized female athymic nude mice (BALB/c/Nu/Nu) were subcutaneously inoculated with MCF-7 cells at the age of 4 weeks. Treatment of mice with established tumors by an intraperitoneal injection of aptamers at doses equivalent to approximately 3 OD/mice/day daily for the first 5 days and an additional treatment at day 7 for a total of 6 injections was performed. The tumor volume was calculated using the formula (1/2 × L × W 2 ). Above, the tumor measurement was made after the mice were sacrificed. The results are shown as a mean of six separate experiments with SD. (C) The body weights of the mice were measured daily, and the results are shown as a mean of six separate experiments with SD.
    Figure Legend Snippet: Modulation of MicroRNA Expression by FCL-II and Suppression Effect of FCL-I/II on MCF-7 Xenografts (A) For TaqMan PCR, the total RNA was isolated using the mirVana miRNA Isolation Kit (Life Technologies Corporation, USA). RT-PCR was performed using the TaqMan MicroRNA Assay (Life Technologies Corporation, USA) and the Mx3005P QPCR System (Agilent Technologies, Inc., CA, USA). U6 gene quantification was used as the control. The results are shown as a mean of three separate experiments with SD. (B) Bilaterally ovariectomized female athymic nude mice (BALB/c/Nu/Nu) were subcutaneously inoculated with MCF-7 cells at the age of 4 weeks. Treatment of mice with established tumors by an intraperitoneal injection of aptamers at doses equivalent to approximately 3 OD/mice/day daily for the first 5 days and an additional treatment at day 7 for a total of 6 injections was performed. The tumor volume was calculated using the formula (1/2 × L × W 2 ). Above, the tumor measurement was made after the mice were sacrificed. The results are shown as a mean of six separate experiments with SD. (C) The body weights of the mice were measured daily, and the results are shown as a mean of six separate experiments with SD.

    Techniques Used: Expressing, Polymerase Chain Reaction, Isolation, Reverse Transcription Polymerase Chain Reaction, TaqMan microRNA Assay, Real-time Polymerase Chain Reaction, Mouse Assay, Injection

    3) Product Images from "Profiling of 95 MicroRNAs in Pancreatic Cancer Cell Lines and Surgical Specimens by Real Time PCR Analysis"

    Article Title: Profiling of 95 MicroRNAs in Pancreatic Cancer Cell Lines and Surgical Specimens by Real Time PCR Analysis

    Journal:

    doi: 10.1007/s00268-008-9833-0

    The expression pattern of 95 miRNAs in chronic pancreatitis, pancreatic cancer cell lines and surgical specimens. MiRNAs of tissues and cultured cells were extracted and purified using mirVana miRNA Isolation kit and converted to cDNAs with the QuantiMir™
    Figure Legend Snippet: The expression pattern of 95 miRNAs in chronic pancreatitis, pancreatic cancer cell lines and surgical specimens. MiRNAs of tissues and cultured cells were extracted and purified using mirVana miRNA Isolation kit and converted to cDNAs with the QuantiMir™

    Techniques Used: Expressing, Cell Culture, Purification, Isolation

    4) Product Images from "Target-dependent biogenesis of cognate microRNAs in human cells"

    Article Title: Target-dependent biogenesis of cognate microRNAs in human cells

    Journal: Nature Communications

    doi: 10.1038/ncomms12200

    Reversal of amino acid-starvation-induced stress increases miR-122 biogenesis in Huh7 cells. ( a , b ) Outline of the experimental set-up used ( a ). Relative level of CAT-1 mRNA in Huh7 cells either fed or starved for 4 h and re-fed with media containing amino acids for another 2 h. CAT-1 mRNA levels were quantified by quantitative reverse transcriptase-PCR with levels in Fed cells taken as 1 ( b ). ( c ) Effect of starvation and re-feeding on mature miR-122 levels in Huh7 cells. Total RNA was extracted and 8 μg RNA was used for northern blotting of mature miR-122, let-7a and miR-16. U6 snRNA was used as loading control. ( d ) Copy number/number of molecules of mature miR-122 and CAT-1 mRNA per Huh7 cell calculated in fed, starved and re-fed Huh7 cells. Estimations were done by real-time-based methods. ( e ) Increase in mature miR-122 but not that of other non-relevant miRNAs, miR-16, miR-21, miR-24 and miR-125b on relief of starvation. Real-time PCR-based quantification of mature miRNA levels in Huh7 cells starved for amino acids (4 h) and subsequently re-fed (2 h). ( f ) Increase in mature miR-122 is accompanied by a concomitant decrease in pre-miR-122 on re-feeding the starved cells for 2 h. Cellular small RNA population was isolated by mirVANA kit to minimize possible contamination of pri-miR-122 and real-time-based assays were carried out to quantify pre-miR-122. Pre-miR-122 detected by northern blotting with 15 μg total RNA. Synthetic pre-miR-122 was used as a size marker to determine the position of the pre-miR-122 in the northern blotting. ( g ) Increase of mature miR-122 on re-feeding of starved cells is reduced in DICER1 knockdown Huh7 cells. miR-16 and miR-21 did not show any significant change. siDICER1-mediated knockdown of DICER1 is confirmed by western blotting. Paired two-tailed Student's t -tests were used for all comparisons. * P
    Figure Legend Snippet: Reversal of amino acid-starvation-induced stress increases miR-122 biogenesis in Huh7 cells. ( a , b ) Outline of the experimental set-up used ( a ). Relative level of CAT-1 mRNA in Huh7 cells either fed or starved for 4 h and re-fed with media containing amino acids for another 2 h. CAT-1 mRNA levels were quantified by quantitative reverse transcriptase-PCR with levels in Fed cells taken as 1 ( b ). ( c ) Effect of starvation and re-feeding on mature miR-122 levels in Huh7 cells. Total RNA was extracted and 8 μg RNA was used for northern blotting of mature miR-122, let-7a and miR-16. U6 snRNA was used as loading control. ( d ) Copy number/number of molecules of mature miR-122 and CAT-1 mRNA per Huh7 cell calculated in fed, starved and re-fed Huh7 cells. Estimations were done by real-time-based methods. ( e ) Increase in mature miR-122 but not that of other non-relevant miRNAs, miR-16, miR-21, miR-24 and miR-125b on relief of starvation. Real-time PCR-based quantification of mature miRNA levels in Huh7 cells starved for amino acids (4 h) and subsequently re-fed (2 h). ( f ) Increase in mature miR-122 is accompanied by a concomitant decrease in pre-miR-122 on re-feeding the starved cells for 2 h. Cellular small RNA population was isolated by mirVANA kit to minimize possible contamination of pri-miR-122 and real-time-based assays were carried out to quantify pre-miR-122. Pre-miR-122 detected by northern blotting with 15 μg total RNA. Synthetic pre-miR-122 was used as a size marker to determine the position of the pre-miR-122 in the northern blotting. ( g ) Increase of mature miR-122 on re-feeding of starved cells is reduced in DICER1 knockdown Huh7 cells. miR-16 and miR-21 did not show any significant change. siDICER1-mediated knockdown of DICER1 is confirmed by western blotting. Paired two-tailed Student's t -tests were used for all comparisons. * P

    Techniques Used: Polymerase Chain Reaction, Northern Blot, Real-time Polymerase Chain Reaction, Isolation, Marker, Western Blot, Two Tailed Test

    5) Product Images from "MicroRNAs Overexpressed in Growth-Restricted Rat Skeletal Muscles Regulate the Glucose Transport in Cell Culture Targeting Central TGF-? Factor SMAD4"

    Article Title: MicroRNAs Overexpressed in Growth-Restricted Rat Skeletal Muscles Regulate the Glucose Transport in Cell Culture Targeting Central TGF-? Factor SMAD4

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0034596

    Semi-quantitative RT-PCR analysis of microRNAs in male SKM samples. I. The total RNA (1 µg) has been used to make cDNA using Ambion mirVana isolation kit. Equal amount of cDNA has been used in separate microRNA-specific PCR reactions. CMCP (1-6) and SMSP (1-6) correspond to six samples of control and starved SKM. II. Image J analyses of the intensities of all bands were plotted as graphs for each microRNAs. T-test was done as a measure of statistical analysis. GAPDH was used as an internal control.
    Figure Legend Snippet: Semi-quantitative RT-PCR analysis of microRNAs in male SKM samples. I. The total RNA (1 µg) has been used to make cDNA using Ambion mirVana isolation kit. Equal amount of cDNA has been used in separate microRNA-specific PCR reactions. CMCP (1-6) and SMSP (1-6) correspond to six samples of control and starved SKM. II. Image J analyses of the intensities of all bands were plotted as graphs for each microRNAs. T-test was done as a measure of statistical analysis. GAPDH was used as an internal control.

    Techniques Used: Quantitative RT-PCR, Isolation, Polymerase Chain Reaction

    6) Product Images from "MicroRNA-186 targets Yes-associated protein 1 to inhibit Hippo signaling and tumorigenesis in hepatocellular carcinoma"

    Article Title: MicroRNA-186 targets Yes-associated protein 1 to inhibit Hippo signaling and tumorigenesis in hepatocellular carcinoma

    Journal:

    doi: 10.3892/ol.2016.4312

    Transfection of miR-186 inhibits hepatocellular carcinoma (HCC) cell migration, invasion and proliferation. (A) Wound-healing assay for three human HCC cell lines (HepG2, Hep3B and SNU398) following transfection with either mirVana miRNA mimic negative
    Figure Legend Snippet: Transfection of miR-186 inhibits hepatocellular carcinoma (HCC) cell migration, invasion and proliferation. (A) Wound-healing assay for three human HCC cell lines (HepG2, Hep3B and SNU398) following transfection with either mirVana miRNA mimic negative

    Techniques Used: Transfection, Migration, Wound Healing Assay

    7) Product Images from "Regulation of chemotropic guidance of nerve growth cones by microRNA"

    Article Title: Regulation of chemotropic guidance of nerve growth cones by microRNA

    Journal: Molecular Brain

    doi: 10.1186/1756-6606-4-40

    Presence of miR-134 in Xenopus neural tissues . ( A ) Taqman stem-loop real-time PCR assays on the expression of three miRNAs, miR-103, miR-134, and miR-191 in rat brain, whole Xenopus embryos, and Xenopus neural tube tissues. Please note that both Xenopus samples were total RNAs, whereas the rat brain sample was processed for microRNA enrichment using a mirVana miRNA isolation kit (see Methods). As a result, the microRNA level in the rat brain sample was much higher and merely serves as a positive control. The relative miRNA levels are plotted in logarithmic scale. The error bars represent the standard error of the mean. Asterisk: p
    Figure Legend Snippet: Presence of miR-134 in Xenopus neural tissues . ( A ) Taqman stem-loop real-time PCR assays on the expression of three miRNAs, miR-103, miR-134, and miR-191 in rat brain, whole Xenopus embryos, and Xenopus neural tube tissues. Please note that both Xenopus samples were total RNAs, whereas the rat brain sample was processed for microRNA enrichment using a mirVana miRNA isolation kit (see Methods). As a result, the microRNA level in the rat brain sample was much higher and merely serves as a positive control. The relative miRNA levels are plotted in logarithmic scale. The error bars represent the standard error of the mean. Asterisk: p

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Isolation, Positive Control

    Related Articles

    Clone Assay:

    Article Title: Mitochondrial translation requires folate-dependent tRNA methylation
    Article Snippet: HCT116 or HEK293T cell lines and sub clones were grown to 70-85% confluency and harvested for mitochondrial extraction (1-2 × 108 ) . .. Mitochondrial tRNAs were extracted using the MirVana™ miRNA Isolation kit (Ambion) for isolation of small RNAs followed by 10% TBE-urea gel purification and extraction as described above (tRNA fraction 65-85 bp).

    Article Title: Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs
    Article Snippet: All animal protocols were approved by the SBMRI Animal Care & Use Committee. .. Total RNA from JK#2 & 11 and SW#5, 6, & 7 clones was prepared using mirVana miRNA Isolation Kit (Ambion) from 5–10 EBs or Puromycin-selected CSs after respective treatments. .. Complementary DNA (cDNA) was synthesized by the QuantiTect RT kit (Qiagen).

    Centrifugation:

    Article Title: Mitochondrial translation requires folate-dependent tRNA methylation
    Article Snippet: Mitochondrial tRNAs were extracted using the MirVana™ miRNA Isolation kit (Ambion) for isolation of small RNAs followed by 10% TBE-urea gel purification and extraction as described above (tRNA fraction 65-85 bp). .. Quantitative analysis of 5-taurinomethyluridine monophosphate (tm5U), 5-taurinomethy-2-thiouridine monophosphate (tm5s2U), 2-thiouridine monophosphate (s2U) and 5-formylcytidine monophosphate (f5C) was performed using high resolution mass spectrometry following a protocol adapted from , .

    Amplification:

    Article Title: Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs
    Article Snippet: Total RNA from JK#2 & 11 and SW#5, 6, & 7 clones was prepared using mirVana miRNA Isolation Kit (Ambion) from 5–10 EBs or Puromycin-selected CSs after respective treatments. .. Real time PCR cycle was 40 cycles (95°C for 20 seconds, 60°C for 30 seconds, and 72°C for 30 seconds).

    Mass Spectrometry:

    Article Title: Mitochondrial translation requires folate-dependent tRNA methylation
    Article Snippet: Mitochondrial tRNAs were extracted using the MirVana™ miRNA Isolation kit (Ambion) for isolation of small RNAs followed by 10% TBE-urea gel purification and extraction as described above (tRNA fraction 65-85 bp). .. Quantitative analysis of 5-taurinomethyluridine monophosphate (tm5U), 5-taurinomethy-2-thiouridine monophosphate (tm5s2U), 2-thiouridine monophosphate (s2U) and 5-formylcytidine monophosphate (f5C) was performed using high resolution mass spectrometry following a protocol adapted from , .

    Positive Control:

    Article Title: Regulation of chemotropic guidance of nerve growth cones by microRNA
    Article Snippet: After inactivation of HRP with 3% H2 O2 , Xlimk1 signals were detected following the same procedure with anti-digoxigenin-HRP (Roche) followed by amplification with Cy3-TSA. .. MirVana™ miRNA Isolation Kit (Applied Biosystems) was used to isolate small RNAs enriched in microRNAs from rat brain, which is used as the positive control for PCR analysis. .. Total RNAs were collected from stage 20-22 Xenopus whole embryos or neural tube tissues.

    Synthesized:

    Article Title: Regulation of chemotropic guidance of nerve growth cones by microRNA
    Article Snippet: MirVana™ miRNA Isolation Kit (Applied Biosystems) was used to isolate small RNAs enriched in microRNAs from rat brain, which is used as the positive control for PCR analysis. .. Total RNAs were collected from stage 20-22 Xenopus whole embryos or neural tube tissues.

    Quantitative RT-PCR:

    Article Title: MicroRNA-186 targets Yes-associated protein 1 to inhibit Hippo signaling and tumorigenesis in hepatocellular carcinoma
    Article Snippet: Paragraph title: RNA extraction and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) ... Total mRNA was extracted with the mirVANA miRNA isolation kit (Ambion Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions.

    Article Title: Inhibition of miR-25 Improves Cardiac Contractility in the Failing Heart
    Article Snippet: Paragraph title: Quantitative RT-PCR ... RNA was isolated from mouse tissue using mirVana™ miRNA Isolation kit (Life Technologies) according to manufacturer’s protocol.

    Article Title: Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs
    Article Snippet: Paragraph title: RNA isolation and quantitative RT-PCR ... Total RNA from JK#2 & 11 and SW#5, 6, & 7 clones was prepared using mirVana miRNA Isolation Kit (Ambion) from 5–10 EBs or Puromycin-selected CSs after respective treatments.

    SYBR Green Assay:

    Article Title: Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs
    Article Snippet: Total RNA from JK#2 & 11 and SW#5, 6, & 7 clones was prepared using mirVana miRNA Isolation Kit (Ambion) from 5–10 EBs or Puromycin-selected CSs after respective treatments. .. Complementary DNA (cDNA) was synthesized by the QuantiTect RT kit (Qiagen).

    Microarray:

    Article Title: The Bioactivity of D-/L-Isonucleoside- and 2′-Deoxyinosine-Incorporated Aptamer AS1411s Including DNA Replication/MicroRNA Expression
    Article Snippet: Paragraph title: Microarray and TaqMan PCR ... For TaqMan PCR, the total RNA was isolated using the mirVana miRNA Isolation Kit (Life Technologies Corporation, USA).

    Article Title: Ovarian surface epithelium at the junction area contains cancer-prone stem cell niche
    Article Snippet: Total RNA was isolated using a mirVana miRNA isolation kit (Ambion, Austin, TX). .. Total RNA was isolated using a mirVana miRNA isolation kit (Ambion, Austin, TX).

    Incubation:

    Article Title: Hypothalamic stem cells control aging speed partly through exosomal miRNAs
    Article Snippet: 5) Pull-down assay: Anti-CD81 antibody and isotype control antibody were coated onto Protein G Beads (Life Technologies, 10003D), exosomes were re-suspended in PBS containing 3 mg/ml BSA and incubated with antibody-coated beads overnight with rotation at 4°C, and subsequently beads were washed with the same buffer. .. Exosomal total RNAs and small RNAs were purified by mirVana miRNA Isolation Kit (Life Technologies).

    Infection:

    Article Title: Two Small RNAs Encoded within the First 1.5 Kilobases of the Herpes Simplex Virus Type 1 Latency-Associated Transcript Can Inhibit Productive Infection and Cooperate To Inhibit Apoptosis
    Article Snippet: TG were extracted during latency (at least 30 days after infection). .. sRNAs were prepared from the TG of latently infected mice or transfected cells using the mirVana miRNA isolation kit (Ambion) according to the manufacturer's instructions. sRNA samples were precipitated in 100% ethanol and suspended in 20 μl RNase-free water, which is referred to as the total sRNA fraction. .. LAT sRNA1 and sRNA2 in TG or transfected cells were detected using a previously described protocol ( ).

    Expressing:

    Article Title: Regulation of chemotropic guidance of nerve growth cones by microRNA
    Article Snippet: Paragraph title: MicroRNA expression analysis ... MirVana™ miRNA Isolation Kit (Applied Biosystems) was used to isolate small RNAs enriched in microRNAs from rat brain, which is used as the positive control for PCR analysis.

    Article Title: Ovarian surface epithelium at the junction area contains cancer-prone stem cell niche
    Article Snippet: Paragraph title: Gene expression arrays ... Total RNA was isolated using a mirVana miRNA isolation kit (Ambion, Austin, TX).

    Article Title: IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity
    Article Snippet: CD44hi CD62Llo CD4+ T cells were sorted from the spleen of naïve Xbp1 f/f or Xbp1 f/f Cd4 cre female mice. miRVana miRNA isolation kit (Ambion) and RNeasy MinElute kit (Qiagen) were used for total RNA isolation and concentration. .. CD44hi CD62Llo CD4+ T cells were sorted from the spleen of naïve Xbp1 f/f or Xbp1 f/f Cd4 cre female mice. miRVana miRNA isolation kit (Ambion) and RNeasy MinElute kit (Qiagen) were used for total RNA isolation and concentration.

    Gel Purification:

    Article Title: Mitochondrial translation requires folate-dependent tRNA methylation
    Article Snippet: HCT116 or HEK293T cell lines and sub clones were grown to 70-85% confluency and harvested for mitochondrial extraction (1-2 × 108 ) . .. Mitochondrial tRNAs were extracted using the MirVana™ miRNA Isolation kit (Ambion) for isolation of small RNAs followed by 10% TBE-urea gel purification and extraction as described above (tRNA fraction 65-85 bp). .. Quantitative analysis of 5-taurinomethyluridine monophosphate (tm5U), 5-taurinomethy-2-thiouridine monophosphate (tm5s2U), 2-thiouridine monophosphate (s2U) and 5-formylcytidine monophosphate (f5C) was performed using high resolution mass spectrometry following a protocol adapted from , .

    Transfection:

    Article Title: Two Small RNAs Encoded within the First 1.5 Kilobases of the Herpes Simplex Virus Type 1 Latency-Associated Transcript Can Inhibit Productive Infection and Cooperate To Inhibit Apoptosis
    Article Snippet: TG were extracted during latency (at least 30 days after infection). .. sRNAs were prepared from the TG of latently infected mice or transfected cells using the mirVana miRNA isolation kit (Ambion) according to the manufacturer's instructions. sRNA samples were precipitated in 100% ethanol and suspended in 20 μl RNase-free water, which is referred to as the total sRNA fraction. .. LAT sRNA1 and sRNA2 in TG or transfected cells were detected using a previously described protocol ( ).

    Northern Blot:

    Article Title: Target-dependent biogenesis of cognate microRNAs in human cells
    Article Snippet: Paragraph title: RNA isolation and northern blotting ... Small RNA was isolated using mirVANA miRNA isolation kit (Life Technologies) according to the manufacturer's instructions.

    Article Title: RNAi triggered by specialized machinery silences developmental genes and retrotransposons
    Article Snippet: Paragraph title: Detection of small RNAs by Northern blot ... For small RNA detection, short RNAs ( < 200nt) were purified with the mirVana miRNA isolation kit (Ambion).

    Article Title: RNAi triggered by specialized machinery silences developmental genes and retrotransposons
    Article Snippet: The preparation of small RNA libraries is described in Full Methods. .. For detection of small RNAs by Northern, 10μg of small RNAs, purified using the mirVana miRNA isolation kit (Ambion), were resolved on a 15% denaturing gel, crosslinked to neutral nylon membranes (Amersham/Pharmacia) and detected using a ~50nt 32 P-labeled single stranded RNA probe. .. Standard protocols were used for cell culture, sporulation and tetrad analysis.

    Article Title: Unique roles for histone H3K9me states in RNAi and heritable silencing of transcription
    Article Snippet: Paragraph title: Small RNA (sRNA) Extraction and Northern Blot ... Briefly, total sRNA was purified with the mirVana miRNA Isolation Kit (Ambion), loaded on 17.5% polyacrylamide/7M urea gel, transferred to positively charged nylon membrane (Roche 1417240), and probed with a mixture of 32 P-labeled DNA probes corresponding to dg or dh siRNA sequences described previously .

    Article Title: Coordination of DNA Replication and Histone Modification by the Rik1-Dos2 Complex
    Article Snippet: Paragraph title: Small RNA Northern Blot ... Briefly, siRNAs were extracted from exponentially growing cells in YES media using a mirVana miRNA isolation kit (Ambion).

    Cell Culture:

    Article Title: Profiling of 95 MicroRNAs in Pancreatic Cancer Cell Lines and Surgical Specimens by Real Time PCR Analysis
    Article Snippet: Human pancreatic adenocarcinoma specimens and their adjacent normal pancreatic tissues (17 pairs) and one pancreatic tissue sample with chronic pancreatitis were collected from patients who underwent surgery according to an approved human protocol at the Baylor College of Medicine (Houston, TX). .. Total miRNAs of tissues and cultured cells were extracted and purified using mirVana miRNA Isolation kit (Applied Biosystems/Ambion, Austin, TX) following the manufacturer's instructions. .. Five µl of RNA was directly converted to cDNA with the QuantiMir™ RT System (SBI System Biosciences, Mountain View, CA).

    Polymerase Chain Reaction:

    Article Title: Regulation of chemotropic guidance of nerve growth cones by microRNA
    Article Snippet: After inactivation of HRP with 3% H2 O2 , Xlimk1 signals were detected following the same procedure with anti-digoxigenin-HRP (Roche) followed by amplification with Cy3-TSA. .. MirVana™ miRNA Isolation Kit (Applied Biosystems) was used to isolate small RNAs enriched in microRNAs from rat brain, which is used as the positive control for PCR analysis. .. Total RNAs were collected from stage 20-22 Xenopus whole embryos or neural tube tissues.

    Article Title: MicroRNA-186 targets Yes-associated protein 1 to inhibit Hippo signaling and tumorigenesis in hepatocellular carcinoma
    Article Snippet: Paragraph title: RNA extraction and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) ... Total mRNA was extracted with the mirVANA miRNA isolation kit (Ambion Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions.

    Article Title: The Bioactivity of D-/L-Isonucleoside- and 2′-Deoxyinosine-Incorporated Aptamer AS1411s Including DNA Replication/MicroRNA Expression
    Article Snippet: RNA preparation and microarray hybridization were performed according to Exiqon’s manual. .. For TaqMan PCR, the total RNA was isolated using the mirVana miRNA Isolation Kit (Life Technologies Corporation, USA). .. RT-PCR was performed using the TaqMan MicroRNA Assay (Life Technologies Corporation, USA) and Mx3005P QPCR System (Agilent Technologies, Inc., CA, USA).

    Article Title: Epigenetic inheritance mediated by coupling of RNAi and histone H3K9 methylation
    Article Snippet: Pellets were processed using the mirVana™ miRNA Isolation kit (Ambion), and the resulting RNA used for library construction. .. Pellets were processed using the mirVana™ miRNA Isolation kit (Ambion), and the resulting RNA used for library construction.

    Protein Concentration:

    Article Title: Hypothalamic stem cells control aging speed partly through exosomal miRNAs
    Article Snippet: 6) Exosomal total protein and RNA: Exosomal total protein concentration was measured by Pierce™ Coomassie (Bradford) Protein Assay Kit (ThermoFisher). .. Exosomal total RNAs and small RNAs were purified by mirVana miRNA Isolation Kit (Life Technologies).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Two Small RNAs Encoded within the First 1.5 Kilobases of the Herpes Simplex Virus Type 1 Latency-Associated Transcript Can Inhibit Productive Infection and Cooperate To Inhibit Apoptosis
    Article Snippet: Paragraph title: Preparation of sRNAs and reverse transcription-PCR (RT-PCR). ... sRNAs were prepared from the TG of latently infected mice or transfected cells using the mirVana miRNA isolation kit (Ambion) according to the manufacturer's instructions. sRNA samples were precipitated in 100% ethanol and suspended in 20 μl RNase-free water, which is referred to as the total sRNA fraction.

    Injection:

    Article Title: Mitochondrial translation requires folate-dependent tRNA methylation
    Article Snippet: Mitochondrial tRNAs were extracted using the MirVana™ miRNA Isolation kit (Ambion) for isolation of small RNAs followed by 10% TBE-urea gel purification and extraction as described above (tRNA fraction 65-85 bp). .. Quantitative analysis of 5-taurinomethyluridine monophosphate (tm5U), 5-taurinomethy-2-thiouridine monophosphate (tm5s2U), 2-thiouridine monophosphate (s2U) and 5-formylcytidine monophosphate (f5C) was performed using high resolution mass spectrometry following a protocol adapted from , .

    Hybridization:

    Article Title: The Bioactivity of D-/L-Isonucleoside- and 2′-Deoxyinosine-Incorporated Aptamer AS1411s Including DNA Replication/MicroRNA Expression
    Article Snippet: RNA preparation and microarray hybridization were performed according to Exiqon’s manual. .. For TaqMan PCR, the total RNA was isolated using the mirVana miRNA Isolation Kit (Life Technologies Corporation, USA).

    Coomassie Assay:

    Article Title: Hypothalamic stem cells control aging speed partly through exosomal miRNAs
    Article Snippet: 6) Exosomal total protein and RNA: Exosomal total protein concentration was measured by Pierce™ Coomassie (Bradford) Protein Assay Kit (ThermoFisher). .. Exosomal total RNAs and small RNAs were purified by mirVana miRNA Isolation Kit (Life Technologies).

    RNA Sequencing Assay:

    Article Title: IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity
    Article Snippet: Paragraph title: RNA-seq and transcriptional profiling ... CD44hi CD62Llo CD4+ T cells were sorted from the spleen of naïve Xbp1 f/f or Xbp1 f/f Cd4 cre female mice. miRVana miRNA isolation kit (Ambion) and RNeasy MinElute kit (Qiagen) were used for total RNA isolation and concentration.

    Pull Down Assay:

    Article Title: Hypothalamic stem cells control aging speed partly through exosomal miRNAs
    Article Snippet: 5) Pull-down assay: Anti-CD81 antibody and isotype control antibody were coated onto Protein G Beads (Life Technologies, 10003D), exosomes were re-suspended in PBS containing 3 mg/ml BSA and incubated with antibody-coated beads overnight with rotation at 4°C, and subsequently beads were washed with the same buffer. .. Exosomal total RNAs and small RNAs were purified by mirVana miRNA Isolation Kit (Life Technologies).

    Fluorescence:

    Article Title: Regulation of chemotropic guidance of nerve growth cones by microRNA
    Article Snippet: MirVana™ miRNA Isolation Kit (Applied Biosystems) was used to isolate small RNAs enriched in microRNAs from rat brain, which is used as the positive control for PCR analysis. .. The first strand cDNA for PCR were synthesized using SuperScript™ III First-Strand Synthesis System (Invitrogen).

    Isolation:

    Article Title: Regulation of chemotropic guidance of nerve growth cones by microRNA
    Article Snippet: After inactivation of HRP with 3% H2 O2 , Xlimk1 signals were detected following the same procedure with anti-digoxigenin-HRP (Roche) followed by amplification with Cy3-TSA. .. MirVana™ miRNA Isolation Kit (Applied Biosystems) was used to isolate small RNAs enriched in microRNAs from rat brain, which is used as the positive control for PCR analysis. .. Total RNAs were collected from stage 20-22 Xenopus whole embryos or neural tube tissues.

    Article Title: Target-dependent biogenesis of cognate microRNAs in human cells
    Article Snippet: Total RNA was isolated using TRIzol reagent (Life Technologies). .. Small RNA was isolated using mirVANA miRNA isolation kit (Life Technologies) according to the manufacturer's instructions. .. Northern blotting of total cellular RNA (5–10 μg) was performed as described by Pillai et al .

    Article Title: MicroRNA-186 targets Yes-associated protein 1 to inhibit Hippo signaling and tumorigenesis in hepatocellular carcinoma
    Article Snippet: Data are shown as the means ± standard error of the mean (SEM) of three independent experiments. .. Total mRNA was extracted with the mirVANA miRNA isolation kit (Ambion Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions. .. The expression level of miR-186 was quantified using miRNA-specific TaqMan Pri-miRNA assays (assay ID Hs03303293_pri; Applied Biosystems) and normalized by RNU48 control miRNA assay (P/N 001006; Applied Biosystems).

    Article Title: The Bioactivity of D-/L-Isonucleoside- and 2′-Deoxyinosine-Incorporated Aptamer AS1411s Including DNA Replication/MicroRNA Expression
    Article Snippet: RNA preparation and microarray hybridization were performed according to Exiqon’s manual. .. For TaqMan PCR, the total RNA was isolated using the mirVana miRNA Isolation Kit (Life Technologies Corporation, USA). .. RT-PCR was performed using the TaqMan MicroRNA Assay (Life Technologies Corporation, USA) and Mx3005P QPCR System (Agilent Technologies, Inc., CA, USA).

    Article Title: Two Small RNAs Encoded within the First 1.5 Kilobases of the Herpes Simplex Virus Type 1 Latency-Associated Transcript Can Inhibit Productive Infection and Cooperate To Inhibit Apoptosis
    Article Snippet: TG were extracted during latency (at least 30 days after infection). .. sRNAs were prepared from the TG of latently infected mice or transfected cells using the mirVana miRNA isolation kit (Ambion) according to the manufacturer's instructions. sRNA samples were precipitated in 100% ethanol and suspended in 20 μl RNase-free water, which is referred to as the total sRNA fraction. .. LAT sRNA1 and sRNA2 in TG or transfected cells were detected using a previously described protocol ( ).

    Article Title: MicroRNAs Overexpressed in Growth-Restricted Rat Skeletal Muscles Regulate the Glucose Transport in Cell Culture Targeting Central TGF-? Factor SMAD4
    Article Snippet: The muscle tissues were cut into pieces and frozen in liquid nitrogen, then ground into powder in mortar and pestles and stored at −80°C. .. For enriched fraction of micro-RNA isolation quantitatively from tissues and cells, the mirVana miRNA isolation kit of Ambion was used. .. RNA quality was measured by mass spectrometry and agarose-gel electrophoresis by examination of the band integrity of ribosomal RNA(s).

    Article Title: Profiling of 95 MicroRNAs in Pancreatic Cancer Cell Lines and Surgical Specimens by Real Time PCR Analysis
    Article Snippet: Human pancreatic adenocarcinoma specimens and their adjacent normal pancreatic tissues (17 pairs) and one pancreatic tissue sample with chronic pancreatitis were collected from patients who underwent surgery according to an approved human protocol at the Baylor College of Medicine (Houston, TX). .. Total miRNAs of tissues and cultured cells were extracted and purified using mirVana miRNA Isolation kit (Applied Biosystems/Ambion, Austin, TX) following the manufacturer's instructions. .. Five µl of RNA was directly converted to cDNA with the QuantiMir™ RT System (SBI System Biosciences, Mountain View, CA).

    Article Title: Inhibition of miR-25 Improves Cardiac Contractility in the Failing Heart
    Article Snippet: Rnu6 was used for normalization. .. RNA was isolated from mouse tissue using mirVana™ miRNA Isolation kit (Life Technologies) according to manufacturer’s protocol. .. The TaqMan® MicroRNA Assays kit (Life Technologies) was used for both cDNA synthesis and quantitative PCR using miR-25 primers (Life Technologies). miR-24 was used for normalization.

    Article Title: Hypothalamic stem cells control aging speed partly through exosomal miRNAs
    Article Snippet: 6) Exosomal total protein and RNA: Exosomal total protein concentration was measured by Pierce™ Coomassie (Bradford) Protein Assay Kit (ThermoFisher). .. Exosomal total RNAs and small RNAs were purified by mirVana miRNA Isolation Kit (Life Technologies). .. Exosomal total RNAs were measured by Qubit assay (ThermoFisher).

    Article Title: Mitochondrial translation requires folate-dependent tRNA methylation
    Article Snippet: HCT116 or HEK293T cell lines and sub clones were grown to 70-85% confluency and harvested for mitochondrial extraction (1-2 × 108 ) . .. Mitochondrial tRNAs were extracted using the MirVana™ miRNA Isolation kit (Ambion) for isolation of small RNAs followed by 10% TBE-urea gel purification and extraction as described above (tRNA fraction 65-85 bp). .. Quantitative analysis of 5-taurinomethyluridine monophosphate (tm5U), 5-taurinomethy-2-thiouridine monophosphate (tm5s2U), 2-thiouridine monophosphate (s2U) and 5-formylcytidine monophosphate (f5C) was performed using high resolution mass spectrometry following a protocol adapted from , .

    Article Title: Ovarian surface epithelium at the junction area contains cancer-prone stem cell niche
    Article Snippet: After 24 hrs adherent cultures hilum and anterior ovary part cells were collected and seeded out for the cell proliferation or OSE-sphere-rim-assays. .. Total RNA was isolated using a mirVana miRNA isolation kit (Ambion, Austin, TX). .. Ovation Pico WTA System V2 (NuGen, San Carlos, CA) was used to amplify cDNA, and then labeled with biotin using Encore Biotin Module (NuGen).

    Article Title: IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity
    Article Snippet: Tumor infiltrating CD4+ T cells (CD45+ CD3+ CD4+ ) were sorted from peritoneal wash samples of Xbp1 f/f or Xbp1 f/f Cd4 cre female mice bearing aggressive ID8-Defb29 /Vegf-A ovarian cancer for 20 days. .. CD44hi CD62Llo CD4+ T cells were sorted from the spleen of naïve Xbp1 f/f or Xbp1 f/f Cd4 cre female mice. miRVana miRNA isolation kit (Ambion) and RNeasy MinElute kit (Qiagen) were used for total RNA isolation and concentration. .. All samples were passed RNA quality controls examined by Agilent Bioanalyzer 2100, and mRNA libraries were generated and sequenced at the Weill Cornell Epigenomics Core Facility.

    Article Title: Antifungal drug resistance evokedvia RNAi-dependent epimutations
    Article Snippet: Sequences were aligned with Serial-Cloner software. .. sRNA libraries were prepared following the instructions of the TruSeq Small RNA Sample Prep Kit (Illumina). sRNA-enriched samples extracted with the mirVana miRNA isolation kit (Ambion) as previously described were size-fractioned by electrophoresis and the 18-24 nt fraction purified from the gel was used as input. .. The library was sequenced by the High-Throughput Sequencing Facility (HTSF), UNC Chapel Hill, using the Illumina platform (HiSeq 2000).

    Article Title: Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs
    Article Snippet: All animal protocols were approved by the SBMRI Animal Care & Use Committee. .. Total RNA from JK#2 & 11 and SW#5, 6, & 7 clones was prepared using mirVana miRNA Isolation Kit (Ambion) from 5–10 EBs or Puromycin-selected CSs after respective treatments. .. Complementary DNA (cDNA) was synthesized by the QuantiTect RT kit (Qiagen).

    Article Title: RNAi triggered by specialized machinery silences developmental genes and retrotransposons
    Article Snippet: The 32 P-labeled single stranded RNA probes were hybridized overnight to the membrane in ULTRAhyb buffer (Ambion). .. For small RNA detection, short RNAs ( < 200nt) were purified with the mirVana miRNA isolation kit (Ambion). .. 10 μg of small RNA was resolved on a 15% denaturing acrylamide gel, followed by transfer to a neutral nylon membrane (Amersham/Pharmacia).

    Article Title: RNAi triggered by specialized machinery silences developmental genes and retrotransposons
    Article Snippet: The preparation of small RNA libraries is described in Full Methods. .. For detection of small RNAs by Northern, 10μg of small RNAs, purified using the mirVana miRNA isolation kit (Ambion), were resolved on a 15% denaturing gel, crosslinked to neutral nylon membranes (Amersham/Pharmacia) and detected using a ~50nt 32 P-labeled single stranded RNA probe. .. Standard protocols were used for cell culture, sporulation and tetrad analysis.

    Article Title: Unique roles for histone H3K9me states in RNAi and heritable silencing of transcription
    Article Snippet: sRNA was purified from 50 ml of cells (OD = ~1.5) as previously described . .. Briefly, total sRNA was purified with the mirVana miRNA Isolation Kit (Ambion), loaded on 17.5% polyacrylamide/7M urea gel, transferred to positively charged nylon membrane (Roche 1417240), and probed with a mixture of 32 P-labeled DNA probes corresponding to dg or dh siRNA sequences described previously . .. Exponentially growing wild-type cells expressing 3xFlag-Clr4 under its endogenous promoter (SPY4636) (2.25 × 1011 cells) were harvested by centrifugation, transferred to a 50 ml tube, washed twice in TBS (50 mM Tris [pH 7.6], 150 mM NaCl), and frozen at −80 °C (approximately 20 g cells).

    Article Title: Coordination of DNA Replication and Histone Modification by the Rik1-Dos2 Complex
    Article Snippet: Small RNA Northern blot was performed as described previously . .. Briefly, siRNAs were extracted from exponentially growing cells in YES media using a mirVana miRNA isolation kit (Ambion). .. 25 μg of small total RNA was resolved by a 15% denaturing acrylamide gel, and blotted to a charged nylon membrane (Hybond-N+, Amersham).

    RNA Extraction:

    Article Title: MicroRNA-186 targets Yes-associated protein 1 to inhibit Hippo signaling and tumorigenesis in hepatocellular carcinoma
    Article Snippet: Paragraph title: RNA extraction and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) ... Total mRNA was extracted with the mirVANA miRNA isolation kit (Ambion Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions.

    Labeling:

    Article Title: Ovarian surface epithelium at the junction area contains cancer-prone stem cell niche
    Article Snippet: Total RNA was isolated using a mirVana miRNA isolation kit (Ambion, Austin, TX). .. Ovation Pico WTA System V2 (NuGen, San Carlos, CA) was used to amplify cDNA, and then labeled with biotin using Encore Biotin Module (NuGen).

    Mouse Assay:

    Article Title: Two Small RNAs Encoded within the First 1.5 Kilobases of the Herpes Simplex Virus Type 1 Latency-Associated Transcript Can Inhibit Productive Infection and Cooperate To Inhibit Apoptosis
    Article Snippet: TG were extracted during latency (at least 30 days after infection). .. sRNAs were prepared from the TG of latently infected mice or transfected cells using the mirVana miRNA isolation kit (Ambion) according to the manufacturer's instructions. sRNA samples were precipitated in 100% ethanol and suspended in 20 μl RNase-free water, which is referred to as the total sRNA fraction. .. LAT sRNA1 and sRNA2 in TG or transfected cells were detected using a previously described protocol ( ).

    Article Title: IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity
    Article Snippet: Tumor infiltrating CD4+ T cells (CD45+ CD3+ CD4+ ) were sorted from peritoneal wash samples of Xbp1 f/f or Xbp1 f/f Cd4 cre female mice bearing aggressive ID8-Defb29 /Vegf-A ovarian cancer for 20 days. .. CD44hi CD62Llo CD4+ T cells were sorted from the spleen of naïve Xbp1 f/f or Xbp1 f/f Cd4 cre female mice. miRVana miRNA isolation kit (Ambion) and RNeasy MinElute kit (Qiagen) were used for total RNA isolation and concentration. .. All samples were passed RNA quality controls examined by Agilent Bioanalyzer 2100, and mRNA libraries were generated and sequenced at the Weill Cornell Epigenomics Core Facility.

    Sequencing:

    Article Title: IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity
    Article Snippet: CD44hi CD62Llo CD4+ T cells were sorted from the spleen of naïve Xbp1 f/f or Xbp1 f/f Cd4 cre female mice. miRVana miRNA isolation kit (Ambion) and RNeasy MinElute kit (Qiagen) were used for total RNA isolation and concentration. .. All samples were passed RNA quality controls examined by Agilent Bioanalyzer 2100, and mRNA libraries were generated and sequenced at the Weill Cornell Epigenomics Core Facility.

    Article Title: Antifungal drug resistance evokedvia RNAi-dependent epimutations
    Article Snippet: Paragraph title: High-throughput sRNA sequencing ... sRNA libraries were prepared following the instructions of the TruSeq Small RNA Sample Prep Kit (Illumina). sRNA-enriched samples extracted with the mirVana miRNA isolation kit (Ambion) as previously described were size-fractioned by electrophoresis and the 18-24 nt fraction purified from the gel was used as input.

    Purification:

    Article Title: MicroRNA-186 targets Yes-associated protein 1 to inhibit Hippo signaling and tumorigenesis in hepatocellular carcinoma
    Article Snippet: Total mRNA was extracted with the mirVANA miRNA isolation kit (Ambion Life Technologies, Carlsbad, CA, USA) according to the manufacturer's instructions. .. The expression level of miR-186 was quantified using miRNA-specific TaqMan Pri-miRNA assays (assay ID Hs03303293_pri; Applied Biosystems) and normalized by RNU48 control miRNA assay (P/N 001006; Applied Biosystems).

    Article Title: Profiling of 95 MicroRNAs in Pancreatic Cancer Cell Lines and Surgical Specimens by Real Time PCR Analysis
    Article Snippet: Human pancreatic adenocarcinoma specimens and their adjacent normal pancreatic tissues (17 pairs) and one pancreatic tissue sample with chronic pancreatitis were collected from patients who underwent surgery according to an approved human protocol at the Baylor College of Medicine (Houston, TX). .. Total miRNAs of tissues and cultured cells were extracted and purified using mirVana miRNA Isolation kit (Applied Biosystems/Ambion, Austin, TX) following the manufacturer's instructions. .. Five µl of RNA was directly converted to cDNA with the QuantiMir™ RT System (SBI System Biosciences, Mountain View, CA).

    Article Title: Hypothalamic stem cells control aging speed partly through exosomal miRNAs
    Article Snippet: 6) Exosomal total protein and RNA: Exosomal total protein concentration was measured by Pierce™ Coomassie (Bradford) Protein Assay Kit (ThermoFisher). .. Exosomal total RNAs and small RNAs were purified by mirVana miRNA Isolation Kit (Life Technologies). .. Exosomal total RNAs were measured by Qubit assay (ThermoFisher).

    Article Title: Antifungal drug resistance evokedvia RNAi-dependent epimutations
    Article Snippet: Sequences were aligned with Serial-Cloner software. .. sRNA libraries were prepared following the instructions of the TruSeq Small RNA Sample Prep Kit (Illumina). sRNA-enriched samples extracted with the mirVana miRNA isolation kit (Ambion) as previously described were size-fractioned by electrophoresis and the 18-24 nt fraction purified from the gel was used as input. .. The library was sequenced by the High-Throughput Sequencing Facility (HTSF), UNC Chapel Hill, using the Illumina platform (HiSeq 2000).

    Article Title: RNAi triggered by specialized machinery silences developmental genes and retrotransposons
    Article Snippet: The 32 P-labeled single stranded RNA probes were hybridized overnight to the membrane in ULTRAhyb buffer (Ambion). .. For small RNA detection, short RNAs ( < 200nt) were purified with the mirVana miRNA isolation kit (Ambion). .. 10 μg of small RNA was resolved on a 15% denaturing acrylamide gel, followed by transfer to a neutral nylon membrane (Amersham/Pharmacia).

    Article Title: RNAi triggered by specialized machinery silences developmental genes and retrotransposons
    Article Snippet: The preparation of small RNA libraries is described in Full Methods. .. For detection of small RNAs by Northern, 10μg of small RNAs, purified using the mirVana miRNA isolation kit (Ambion), were resolved on a 15% denaturing gel, crosslinked to neutral nylon membranes (Amersham/Pharmacia) and detected using a ~50nt 32 P-labeled single stranded RNA probe. .. Standard protocols were used for cell culture, sporulation and tetrad analysis.

    Article Title: Unique roles for histone H3K9me states in RNAi and heritable silencing of transcription
    Article Snippet: sRNA was purified from 50 ml of cells (OD = ~1.5) as previously described . .. Briefly, total sRNA was purified with the mirVana miRNA Isolation Kit (Ambion), loaded on 17.5% polyacrylamide/7M urea gel, transferred to positively charged nylon membrane (Roche 1417240), and probed with a mixture of 32 P-labeled DNA probes corresponding to dg or dh siRNA sequences described previously . .. Exponentially growing wild-type cells expressing 3xFlag-Clr4 under its endogenous promoter (SPY4636) (2.25 × 1011 cells) were harvested by centrifugation, transferred to a 50 ml tube, washed twice in TBS (50 mM Tris [pH 7.6], 150 mM NaCl), and frozen at −80 °C (approximately 20 g cells).

    Liquid Chromatography:

    Article Title: Mitochondrial translation requires folate-dependent tRNA methylation
    Article Snippet: Mitochondrial tRNAs were extracted using the MirVana™ miRNA Isolation kit (Ambion) for isolation of small RNAs followed by 10% TBE-urea gel purification and extraction as described above (tRNA fraction 65-85 bp). .. Quantitative analysis of 5-taurinomethyluridine monophosphate (tm5U), 5-taurinomethy-2-thiouridine monophosphate (tm5s2U), 2-thiouridine monophosphate (s2U) and 5-formylcytidine monophosphate (f5C) was performed using high resolution mass spectrometry following a protocol adapted from , .

    Software:

    Article Title: Regulation of chemotropic guidance of nerve growth cones by microRNA
    Article Snippet: MirVana™ miRNA Isolation Kit (Applied Biosystems) was used to isolate small RNAs enriched in microRNAs from rat brain, which is used as the positive control for PCR analysis. .. The first strand cDNA for PCR were synthesized using SuperScript™ III First-Strand Synthesis System (Invitrogen).

    Article Title: Ovarian surface epithelium at the junction area contains cancer-prone stem cell niche
    Article Snippet: Total RNA was isolated using a mirVana miRNA isolation kit (Ambion, Austin, TX). .. All microarray analyses were performed as previously described , , , .

    Hydrophilic Interaction Liquid Chromatography:

    Article Title: Mitochondrial translation requires folate-dependent tRNA methylation
    Article Snippet: Mitochondrial tRNAs were extracted using the MirVana™ miRNA Isolation kit (Ambion) for isolation of small RNAs followed by 10% TBE-urea gel purification and extraction as described above (tRNA fraction 65-85 bp). .. In brief, 100 ng tRNA were digested at 37°C for 2 h by nuclease P1 (2 U) in 30 μl of 100 mM ammonium acetate and quenched by adding 60 μl 50/50 methanol/acetonitrile followed by centrifugation at 10, 000 g for 10 min. 5 μl of sample was injected for liquid-chromatography mass spectrometry analysis.

    Real-time Polymerase Chain Reaction:

    Article Title: Regulation of chemotropic guidance of nerve growth cones by microRNA
    Article Snippet: MirVana™ miRNA Isolation Kit (Applied Biosystems) was used to isolate small RNAs enriched in microRNAs from rat brain, which is used as the positive control for PCR analysis. .. The first strand cDNA for PCR were synthesized using SuperScript™ III First-Strand Synthesis System (Invitrogen).

    Article Title: Hypothalamic stem cells control aging speed partly through exosomal miRNAs
    Article Snippet: Isolated exosomes were analyzed by real-time PCR. .. Exosomal total RNAs and small RNAs were purified by mirVana miRNA Isolation Kit (Life Technologies).

    Article Title: Studying arrhythmogenic right ventricular dysplasia with patient-specific iPSCs
    Article Snippet: Total RNA from JK#2 & 11 and SW#5, 6, & 7 clones was prepared using mirVana miRNA Isolation Kit (Ambion) from 5–10 EBs or Puromycin-selected CSs after respective treatments. .. Total RNA from JK#2 & 11 and SW#5, 6, & 7 clones was prepared using mirVana miRNA Isolation Kit (Ambion) from 5–10 EBs or Puromycin-selected CSs after respective treatments.

    Multiplex Assay:

    Article Title: Antifungal drug resistance evokedvia RNAi-dependent epimutations
    Article Snippet: sRNA libraries were prepared following the instructions of the TruSeq Small RNA Sample Prep Kit (Illumina). sRNA-enriched samples extracted with the mirVana miRNA isolation kit (Ambion) as previously described were size-fractioned by electrophoresis and the 18-24 nt fraction purified from the gel was used as input. .. sRNA libraries were prepared following the instructions of the TruSeq Small RNA Sample Prep Kit (Illumina). sRNA-enriched samples extracted with the mirVana miRNA isolation kit (Ambion) as previously described were size-fractioned by electrophoresis and the 18-24 nt fraction purified from the gel was used as input.

    Sample Prep:

    Article Title: Antifungal drug resistance evokedvia RNAi-dependent epimutations
    Article Snippet: Sequences were aligned with Serial-Cloner software. .. sRNA libraries were prepared following the instructions of the TruSeq Small RNA Sample Prep Kit (Illumina). sRNA-enriched samples extracted with the mirVana miRNA isolation kit (Ambion) as previously described were size-fractioned by electrophoresis and the 18-24 nt fraction purified from the gel was used as input. .. The library was sequenced by the High-Throughput Sequencing Facility (HTSF), UNC Chapel Hill, using the Illumina platform (HiSeq 2000).

    Electrophoresis:

    Article Title: Antifungal drug resistance evokedvia RNAi-dependent epimutations
    Article Snippet: Sequences were aligned with Serial-Cloner software. .. sRNA libraries were prepared following the instructions of the TruSeq Small RNA Sample Prep Kit (Illumina). sRNA-enriched samples extracted with the mirVana miRNA isolation kit (Ambion) as previously described were size-fractioned by electrophoresis and the 18-24 nt fraction purified from the gel was used as input. .. The library was sequenced by the High-Throughput Sequencing Facility (HTSF), UNC Chapel Hill, using the Illumina platform (HiSeq 2000).

    Knock-Out:

    Article Title: Mitochondrial translation requires folate-dependent tRNA methylation
    Article Snippet: Mitochondrial tRNAs were extracted using the MirVana™ miRNA Isolation kit (Ambion) for isolation of small RNAs followed by 10% TBE-urea gel purification and extraction as described above (tRNA fraction 65-85 bp). .. Nucleoside monophosphates were analyzed on a quadrupole-orbitrap mass spectrometer (Q Exactive plus, Thermo Fisher Scientific) operating in negative ion mode coupled to hydrophilic interaction chromatography via electrospray ionization and used to scan in SIM mode from m/z 459 to 482 (tm5U and tm5s2U) or m/z 338 to 355 (s2U and f5C) at 1 Hz and 140,000 resolution.

    Produced:

    Article Title: IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity
    Article Snippet: CD44hi CD62Llo CD4+ T cells were sorted from the spleen of naïve Xbp1 f/f or Xbp1 f/f Cd4 cre female mice. miRVana miRNA isolation kit (Ambion) and RNeasy MinElute kit (Qiagen) were used for total RNA isolation and concentration. .. All samples were passed RNA quality controls examined by Agilent Bioanalyzer 2100, and mRNA libraries were generated and sequenced at the Weill Cornell Epigenomics Core Facility.

    Concentration Assay:

    Article Title: The Bioactivity of D-/L-Isonucleoside- and 2′-Deoxyinosine-Incorporated Aptamer AS1411s Including DNA Replication/MicroRNA Expression
    Article Snippet: After 16 hr to allow for the adherence of the cells, oligos (or PBS as control) were directly added to the culture medium at a final concentration of 20 μM. .. For TaqMan PCR, the total RNA was isolated using the mirVana miRNA Isolation Kit (Life Technologies Corporation, USA).

    Article Title: Epigenetic inheritance mediated by coupling of RNAi and histone H3K9 methylation
    Article Snippet: To purify total sRNAs, cells were grown in 20 ml YES to a concentration of ~2×107 cells/ml. .. Pellets were processed using the mirVana™ miRNA Isolation kit (Ambion), and the resulting RNA used for library construction.

    Article Title: IRE1α-XBP1 controls T cell function in ovarian cancer by regulating mitochondrial activity
    Article Snippet: Tumor infiltrating CD4+ T cells (CD45+ CD3+ CD4+ ) were sorted from peritoneal wash samples of Xbp1 f/f or Xbp1 f/f Cd4 cre female mice bearing aggressive ID8-Defb29 /Vegf-A ovarian cancer for 20 days. .. CD44hi CD62Llo CD4+ T cells were sorted from the spleen of naïve Xbp1 f/f or Xbp1 f/f Cd4 cre female mice. miRVana miRNA isolation kit (Ambion) and RNeasy MinElute kit (Qiagen) were used for total RNA isolation and concentration. .. All samples were passed RNA quality controls examined by Agilent Bioanalyzer 2100, and mRNA libraries were generated and sequenced at the Weill Cornell Epigenomics Core Facility.

    High Throughput Screening Assay:

    Article Title: Antifungal drug resistance evokedvia RNAi-dependent epimutations
    Article Snippet: Paragraph title: High-throughput sRNA sequencing ... sRNA libraries were prepared following the instructions of the TruSeq Small RNA Sample Prep Kit (Illumina). sRNA-enriched samples extracted with the mirVana miRNA isolation kit (Ambion) as previously described were size-fractioned by electrophoresis and the 18-24 nt fraction purified from the gel was used as input.

    RNA Detection:

    Article Title: RNAi triggered by specialized machinery silences developmental genes and retrotransposons
    Article Snippet: The 32 P-labeled single stranded RNA probes were hybridized overnight to the membrane in ULTRAhyb buffer (Ambion). .. For small RNA detection, short RNAs ( < 200nt) were purified with the mirVana miRNA isolation kit (Ambion). .. 10 μg of small RNA was resolved on a 15% denaturing acrylamide gel, followed by transfer to a neutral nylon membrane (Amersham/Pharmacia).

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    Thermo Fisher mirvana mirna isolation kit with phenol
    Mirvana Mirna Isolation Kit With Phenol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3111 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 99 stars, based on 3111 article reviews
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    mirvana mirna isolation kit with phenol - by Bioz Stars, 2019-12
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