superscript ii cdna synthesis kit thermo fisher scientific  (Thermo Fisher)


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    Name:
    SuperScript Double Stranded cDNA Synthesis Kit
    Description:
    The SuperScript Double Stranded cDNA Synthesis Kit contains all of the reagents except an oligo dT containing primer necessary to make high quality double stranded cDNA from total RNA or poly A selected RNA mRNA The advanced capabilities of SuperScript II Reverse Transcriptase with its reduced RNase H activity maximize the yield of full length cDNA as well as that of overall cDNA synthesis Features of the SuperScript Double Stranded cDNA Synthesis Kit include • Reliability each buffer reagent and enzyme in the kit is of the same high quality you expect from us• Performance SuperScript II is an enzyme you can trust for high yield high quality general purpose first strand synthesis Simple reliable and trouble freeThe SuperScript Double Stranded cDNA Synthesis Kit offers a simple reliable and trouble free method by which RNA templates can be converted to cDNA for use in cloning and library construction experiments The reactions described in the SuperScript Double Stranded cDNA Synthesis Kit protocol are designed to convert 25 50 µg total RNA or 0 2 5 µg mRNA into first and second strand cDNA supply your own primer Enjoy the benefits of a tested protocol and reagents of consistent quality To get the best possible cDNA product using this kit we recommend the use of a high quality RNA substrate prepared using TRIzol RNA Isolation Reagents
    Catalog Number:
    11917010
    Price:
    None
    Applications:
    DNA & RNA Purification & Analysis|DNA Labeling|Nucleic Acid Labeling & Oligo Synthesis|PCR & Real-Time PCR|Reverse Transcription|Gene Expression Analysis & Genotyping
    Category:
    Kits and Assays
    Buy from Supplier


    Structured Review

    Thermo Fisher superscript ii cdna synthesis kit thermo fisher scientific
    The SuperScript Double Stranded cDNA Synthesis Kit contains all of the reagents except an oligo dT containing primer necessary to make high quality double stranded cDNA from total RNA or poly A selected RNA mRNA The advanced capabilities of SuperScript II Reverse Transcriptase with its reduced RNase H activity maximize the yield of full length cDNA as well as that of overall cDNA synthesis Features of the SuperScript Double Stranded cDNA Synthesis Kit include • Reliability each buffer reagent and enzyme in the kit is of the same high quality you expect from us• Performance SuperScript II is an enzyme you can trust for high yield high quality general purpose first strand synthesis Simple reliable and trouble freeThe SuperScript Double Stranded cDNA Synthesis Kit offers a simple reliable and trouble free method by which RNA templates can be converted to cDNA for use in cloning and library construction experiments The reactions described in the SuperScript Double Stranded cDNA Synthesis Kit protocol are designed to convert 25 50 µg total RNA or 0 2 5 µg mRNA into first and second strand cDNA supply your own primer Enjoy the benefits of a tested protocol and reagents of consistent quality To get the best possible cDNA product using this kit we recommend the use of a high quality RNA substrate prepared using TRIzol RNA Isolation Reagents
    https://www.bioz.com/result/superscript ii cdna synthesis kit thermo fisher scientific/product/Thermo Fisher
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    superscript ii cdna synthesis kit thermo fisher scientific - by Bioz Stars, 2020-01
    90/100 stars

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    Related Articles

    Centrifugation:

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: A final nutrient-replete culture was placed in the dark for 24 h, shifted into full light and then harvested after 2 h. For all harvests, triplicate 50-ml aliquots of culture were collected by centrifugation (1 min, 10 000 g ) and flash frozen in liquid nitrogen. .. OligoDT primers were used for cDNA synthesis (SuperScript III, Life Technologies) followed by the the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for second-strand synthesis.

    Amplification:

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: .. Amplified RNA was converted to double-stranded cDNA using the SuperScript III First-Strand Synthesis System (Life Technologies) with priming via random hexamers for the first-strand synthesis and the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for the second-strand synthesis. cDNA in the 0.3–3.0-kb size range was purified from agarose gels using the QIAquick Gel Extraction Kit reagents and protocols (Qiagen) and digested with BpmI for 2–3 h at 37 °C to remove poly(A) tails. .. Following BpmI digestion, cDNA was purified with Ampure XP beads (Beckman Coulter, Brea, CA, USA) and used directly for pyrosequencing.

    Article Title: Single-molecule long-read sequencing facilitates shrimp transcriptome research
    Article Snippet: Double-stranded cDNAs were synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, USA) with random hexamer primers (Illumina, USA), following the manufacturer’s instructions. .. Synthesized cDNAs were gen-purified and amplified with PCR.

    Article Title: Rsx, a metatherian RNA with Xist-like properties
    Article Snippet: Reaction mixture was purified using QIAquick PCR purification kit (Qiagen) and second strand synthesis was performed using SuperScript double-stranded cDNA synthesis kit (Invitrogen) as recommended by the manufacturer with the exception that 200 µM dTTP was replaced with 400 µM dUTP. .. Before final PCR amplification samples were treated with Uracil-N-Glycosylase (Applied Biosystems).

    Article Title: Experimental Incubations Elicit Profound Changes in Community Transcription in OMZ Bacterioplankton
    Article Snippet: Paragraph title: rRNA subtraction, RNA amplification and cDNA synthesis ... Antisense RNA (∼5–10 µg aliquot) was then converted to double-stranded cDNA using the SuperScript® III First-Strand Synthesis System (Invitrogen) with priming via random hexamers for first-strand synthesis, and the SuperScript™ Double-Stranded cDNA synthesis kit (Invitrogen) for second-strand synthesis. cDNA was then purified with the QIAquick PCR purification kit (Qiagen), restriction-digested with BpmI to remove poly(A) tails, and used directly for pyrosequencing

    Article Title: Pax3 isoforms in sensory neurogenesis: expression and function in the ophthalmic trigeminal placode
    Article Snippet: Reverse transcription of RNA was performed using a Superscript Double-Stranded cDNA Synthesis Kit from Invitrogen with oligo primers. .. Forward and reverse primers used for initial PCR amplification of the splice variants are diagramed in as follows: P1 = 5′-CTGCGTCTCCAAGATCCTCT-3′; P2 = 3′-AAAAGCCATCAGTTGGTTGG-5′; P3 = 5′-AGCAACTGGAAGAGCTGGAA-3′; P4 = 3′-TGACAGGGTCCATGCTGTAG-5′.

    Synthesized:

    Article Title: Leptin Stimulates Cellular Glycolysis Through a STAT3 Dependent Mechanism in Tilapia
    Article Snippet: .. The cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen) and tagged with a 5′, four-nucleotide barcode. .. Illumina cDNA libraries (n = 3) were prepared for each treatment (control or leptin).

    Article Title: Identification and functional clustering of global gene expression differences between human age-related cataract and clear lenses
    Article Snippet: .. First and second strand cDNAs were synthesized from 2–5 μg of total RNA using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen, Gaithersburg, MD) and the oligo-dT24 -T7 primer (5′-GGC CAG TGA ATT GTA ATA CGA CTC ACT AT-AGG GAG GCG G-3′) according to the manufacturer's instructions. cRNA was synthesized and labeled with biotinylated UTP and CTP by in vitro transcription using the T7 promoter coupled double-stranded cDNA as a template and the T7 RNA Transcript Labeling Kit (ENZO Diagnostics Inc., Farmingdale, NY). .. Briefly, double-stranded cDNAs synthesized from the previous steps were washed twice with 70% ethanol and resuspended in 22 μl of RNase-free H2 O.

    Article Title: Transcriptional Profiling of Lipopolysaccharide-Induced Acute Lung Injury
    Article Snippet: .. Single-stranded and then double-stranded cDNAs were synthesized from the poly(A+ ) mRNA present in the purified total RNA by use of a SuperScript double-stranded cDNA synthesis kit (Invitrogen Corp.) and poly(T) nucleotide primers that contained a sequence recognized by T7 RNA polymerase. .. The resulting double-stranded cDNAs were used as templates to generate biotinylated cRNAs by in vitro transcription reactions with a Bio-Array High-Yield RNA transcript labeling kit (Enzo Diagnostics Inc., Farmingdale, N.Y.).

    Article Title: Ab initio construction of a eukaryotic transcriptome by massively parallel mRNA sequencing
    Article Snippet: .. The cDNA was synthesized by using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen) with SuperScript III (Invitrogen), 15-ng random hexamers (Invitrogen), and 20 units SUPERase·In (Ambion). .. Primer annealing was done at room temperature for 10 min followed by 1 h at 55 °C for first strand synthesis and 2 h at 16 °C for second strand synthesis; cDNA was sheared by sonication with 12 alternating cycles between “high intensity” (30 s; duty cycle, 20%; intensity, 10%; cycles per burst, 200) and “low intensity” (4 s; duty cycle, 5%; intensity, 10%; cycles per burst, 200) in the Frequency Sweeping mode (Covaris S2 machine).

    Article Title: Single-molecule long-read sequencing facilitates shrimp transcriptome research
    Article Snippet: .. Double-stranded cDNAs were synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, USA) with random hexamer primers (Illumina, USA), following the manufacturer’s instructions. .. Synthesized cDNAs were gen-purified and amplified with PCR.

    Article Title: De novo assembly and characterization of root transcriptome using Illumina paired-end sequencing and development of cSSR markers in sweetpotato (Ipomoea batatas)
    Article Snippet: .. Then the double-stranded cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen, Camarillo, CA) with random hexamer (N6) primers (Illumina). .. The synthesized cDNA was subjected to end-repair and phosphorylation using T4 DNA polymerase, Klenow DNA polymerase and T4 PNK.

    Article Title: Ab initio construction of a eukaryotic transcriptome by massively parallel mRNA sequencing
    Article Snippet: .. RNA from the HS and YPD reference samples was treated with TURBO DNA-free Kit (Ambion) to remove trace amounts of genomic DNA; cDNA was synthesized from this RNA by using a SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen). .. Assays were designed to detect predicted RNA species by the PCR.

    Article Title: Mining Gene Expression Data for Pollutants (Dioxin, Toluene, Formaldehyde) and Low Dose of Gamma-Irradiation
    Article Snippet: .. Briefly, cDNA was synthesized from fragmented RNA using a SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen, USA). ..

    Construct:

    Article Title: Single-molecule long-read sequencing facilitates shrimp transcriptome research
    Article Snippet: Illumina library construction and sequencing The Illumina libraries used to correct the FLNC reads were constructed with the Tru-Seq RNA sample Prep kit (Illumina, USA). .. Double-stranded cDNAs were synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, USA) with random hexamer primers (Illumina, USA), following the manufacturer’s instructions.

    Microarray:

    Article Title: Identification and functional clustering of global gene expression differences between human age-related cataract and clear lenses
    Article Snippet: Paragraph title: Microarray procedure and analysis ... First and second strand cDNAs were synthesized from 2–5 μg of total RNA using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen, Gaithersburg, MD) and the oligo-dT24 -T7 primer (5′-GGC CAG TGA ATT GTA ATA CGA CTC ACT AT-AGG GAG GCG G-3′) according to the manufacturer's instructions. cRNA was synthesized and labeled with biotinylated UTP and CTP by in vitro transcription using the T7 promoter coupled double-stranded cDNA as a template and the T7 RNA Transcript Labeling Kit (ENZO Diagnostics Inc., Farmingdale, NY).

    Article Title: Transcriptional Profiling of Lipopolysaccharide-Induced Acute Lung Injury
    Article Snippet: A mouse Moe430 microarray set that represents approximately 23,000 mouse genes and expressed sequence tag clusters was used to determine the changes in mRNA expression in response to E. coli LPS. .. Single-stranded and then double-stranded cDNAs were synthesized from the poly(A+ ) mRNA present in the purified total RNA by use of a SuperScript double-stranded cDNA synthesis kit (Invitrogen Corp.) and poly(T) nucleotide primers that contained a sequence recognized by T7 RNA polymerase.

    Article Title: Transcriptional Dysregulation in NIPBL and Cohesin Mutant Human Cells
    Article Snippet: RNA Isolation and Affymatrix Expression Array Hybridization Total RNA from each sample was extracted with the RNeasy Mini-kit (Qiagen), synthesis of double-stranded cDNA was performed using SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen), and cleaned up with GeneChip Sample Cleanup module (Affymetrix). .. 30 µl fragmented cRNA at the concentration of 500 ng/µl was sent for hybridization in the microarray facility at The Children's Hospital of Philadelphia.

    Incubation:

    Article Title: Leptin Stimulates Cellular Glycolysis Through a STAT3 Dependent Mechanism in Tilapia
    Article Snippet: RNA-seq and bioinformatic analyses of the pituitary transcriptome For RNA-seq studies assessing leptin modulation of the pituitary RPD transcriptome, tissues were placed in wells of a Falcon 6-well plate in 3 mL of either hormone-free (control) or leptin-containing medium and incubated for 6 h (n = 30/treatment; 10 RPD/well, 3 wells/treatment). .. The cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen) and tagged with a 5′, four-nucleotide barcode.

    Article Title: Identification and functional clustering of global gene expression differences between human age-related cataract and clear lenses
    Article Snippet: First and second strand cDNAs were synthesized from 2–5 μg of total RNA using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen, Gaithersburg, MD) and the oligo-dT24 -T7 primer (5′-GGC CAG TGA ATT GTA ATA CGA CTC ACT AT-AGG GAG GCG G-3′) according to the manufacturer's instructions. cRNA was synthesized and labeled with biotinylated UTP and CTP by in vitro transcription using the T7 promoter coupled double-stranded cDNA as a template and the T7 RNA Transcript Labeling Kit (ENZO Diagnostics Inc., Farmingdale, NY). .. The cDNA was incubated with 4 μl each of 10X Reaction Buffer, Biotin Labeled Ribonucleotides, DTT, RNase Inhibitor Mix, and 2 μl of 20X T7 RNA Polymerase for 5 h at 37 °C.

    Expressing:

    Article Title: Transcriptional Profiling of Lipopolysaccharide-Induced Acute Lung Injury
    Article Snippet: A mouse Moe430 microarray set that represents approximately 23,000 mouse genes and expressed sequence tag clusters was used to determine the changes in mRNA expression in response to E. coli LPS. .. Single-stranded and then double-stranded cDNAs were synthesized from the poly(A+ ) mRNA present in the purified total RNA by use of a SuperScript double-stranded cDNA synthesis kit (Invitrogen Corp.) and poly(T) nucleotide primers that contained a sequence recognized by T7 RNA polymerase.

    Article Title: Transcriptional Dysregulation in NIPBL and Cohesin Mutant Human Cells
    Article Snippet: .. RNA Isolation and Affymatrix Expression Array Hybridization Total RNA from each sample was extracted with the RNeasy Mini-kit (Qiagen), synthesis of double-stranded cDNA was performed using SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen), and cleaned up with GeneChip Sample Cleanup module (Affymetrix). .. The resulting products were then used to synthesize biotin-labeled cRNA with Enzo Bioarray High Yield RNA Transcript Labeling kit (Enzo Life Sciences) and further fragmented to 35–200-bp oligos.

    Modification:

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: Briefly, amplified rRNA-subtracted RNA was converted to double-stranded cDNA via reverse transcription primed with a modified Oligo(dT) primer containing a promoter sequence for T7 RNA polymerase and a recognition site for the restriction enzyme BpmI ( ). cDNA was then subjected to two rounds of in vitro transcription at 37 °C for 14 h yielding large quantities (100–200 μg) of single-stranded antisense RNA. .. Amplified RNA was converted to double-stranded cDNA using the SuperScript III First-Strand Synthesis System (Life Technologies) with priming via random hexamers for the first-strand synthesis and the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for the second-strand synthesis. cDNA in the 0.3–3.0-kb size range was purified from agarose gels using the QIAquick Gel Extraction Kit reagents and protocols (Qiagen) and digested with BpmI for 2–3 h at 37 °C to remove poly(A) tails.

    Hybridization:

    Article Title: Comparative Transcriptional Profiling of Two Contrasting Barley Genotypes under Salinity Stress during the Seedling Stage
    Article Snippet: Paragraph title: 2.3. RNA Preparations and Gene Chip Hybridization ... The SuperScript double-stranded cDNA synthesis kit (Invitrogen) was used to synthesize complementary DNA.

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: Multiple rounds of subtractive hybridization of rRNAs were used to obtain an average of 30 ng of rRNA-subtracted total RNA. .. Amplified RNA was converted to double-stranded cDNA using the SuperScript III First-Strand Synthesis System (Life Technologies) with priming via random hexamers for the first-strand synthesis and the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for the second-strand synthesis. cDNA in the 0.3–3.0-kb size range was purified from agarose gels using the QIAquick Gel Extraction Kit reagents and protocols (Qiagen) and digested with BpmI for 2–3 h at 37 °C to remove poly(A) tails.

    Article Title: Transcriptional Profiling of Lipopolysaccharide-Induced Acute Lung Injury
    Article Snippet: Single-stranded and then double-stranded cDNAs were synthesized from the poly(A+ ) mRNA present in the purified total RNA by use of a SuperScript double-stranded cDNA synthesis kit (Invitrogen Corp.) and poly(T) nucleotide primers that contained a sequence recognized by T7 RNA polymerase. .. Subsequently, 10 μg of this fragmented cRNA (target) was hybridized at 45°C with rotation for 16 h (Affymetrix GeneChip Hybridization Oven 320) to probe sets present on a Moe430A array.

    Article Title: Transcriptional Dysregulation in NIPBL and Cohesin Mutant Human Cells
    Article Snippet: .. RNA Isolation and Affymatrix Expression Array Hybridization Total RNA from each sample was extracted with the RNeasy Mini-kit (Qiagen), synthesis of double-stranded cDNA was performed using SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen), and cleaned up with GeneChip Sample Cleanup module (Affymetrix). .. The resulting products were then used to synthesize biotin-labeled cRNA with Enzo Bioarray High Yield RNA Transcript Labeling kit (Enzo Life Sciences) and further fragmented to 35–200-bp oligos.

    Article Title: Experimental Incubations Elicit Profound Changes in Community Transcription in OMZ Bacterioplankton
    Article Snippet: rRNA subtraction, RNA amplification and cDNA synthesis Bacterial, Archaeal, and Eukaryal ribosomal RNA transcripts were removed from total RNA extracts via a subtractive hybridization protocol using sample-specific rRNA probes based on amplicons obtained from PCR of rRNA genes from source water community DNA, as described in . rRNA-depleted total RNA was then amplified using the MessageAmp™ II-Bacteria kit (Ambion) as described previously , . .. Antisense RNA (∼5–10 µg aliquot) was then converted to double-stranded cDNA using the SuperScript® III First-Strand Synthesis System (Invitrogen) with priming via random hexamers for first-strand synthesis, and the SuperScript™ Double-Stranded cDNA synthesis kit (Invitrogen) for second-strand synthesis. cDNA was then purified with the QIAquick PCR purification kit (Qiagen), restriction-digested with BpmI to remove poly(A) tails, and used directly for pyrosequencing

    Polymerase Chain Reaction:

    Article Title: Ab initio construction of a eukaryotic transcriptome by massively parallel mRNA sequencing
    Article Snippet: The cDNA was synthesized by using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen) with SuperScript III (Invitrogen), 15-ng random hexamers (Invitrogen), and 20 units SUPERase·In (Ambion). .. Adapters for Illumina sequencing were added following the instructions provided, except that 5 times less adapter mix was ligated to the cDNAs and PCR primers were removed by digestion with RecJ (New England Biolabs).

    Article Title: Single-molecule long-read sequencing facilitates shrimp transcriptome research
    Article Snippet: Double-stranded cDNAs were synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, USA) with random hexamer primers (Illumina, USA), following the manufacturer’s instructions. .. Synthesized cDNAs were gen-purified and amplified with PCR.

    Article Title: Rsx, a metatherian RNA with Xist-like properties
    Article Snippet: .. Reaction mixture was purified using QIAquick PCR purification kit (Qiagen) and second strand synthesis was performed using SuperScript double-stranded cDNA synthesis kit (Invitrogen) as recommended by the manufacturer with the exception that 200 µM dTTP was replaced with 400 µM dUTP. .. Double-stranded cDNA was fragmented using Bioruptor (Diagenode) (15 min, Low power, 30 sec ON, 30 sec OFF) in 50 µl volume.

    Article Title: Ab initio construction of a eukaryotic transcriptome by massively parallel mRNA sequencing
    Article Snippet: RNA from the HS and YPD reference samples was treated with TURBO DNA-free Kit (Ambion) to remove trace amounts of genomic DNA; cDNA was synthesized from this RNA by using a SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen). .. Assays were designed to detect predicted RNA species by the PCR.

    Article Title: Experimental Incubations Elicit Profound Changes in Community Transcription in OMZ Bacterioplankton
    Article Snippet: .. Antisense RNA (∼5–10 µg aliquot) was then converted to double-stranded cDNA using the SuperScript® III First-Strand Synthesis System (Invitrogen) with priming via random hexamers for first-strand synthesis, and the SuperScript™ Double-Stranded cDNA synthesis kit (Invitrogen) for second-strand synthesis. cDNA was then purified with the QIAquick PCR purification kit (Qiagen), restriction-digested with BpmI to remove poly(A) tails, and used directly for pyrosequencing .. Pyrosequencing Using standard protocols (454 Life Sciences, Roche), poly(A)-removed cDNA was purified via the AMPure® kit (Agencourt®) and used to generate single-stranded DNA libraries for use as template in emulsion PCR.

    Article Title: Pax3 isoforms in sensory neurogenesis: expression and function in the ophthalmic trigeminal placode
    Article Snippet: Reverse transcription of RNA was performed using a Superscript Double-Stranded cDNA Synthesis Kit from Invitrogen with oligo primers. .. Forward and reverse primers used for initial PCR amplification of the splice variants are diagramed in as follows: P1 = 5′-CTGCGTCTCCAAGATCCTCT-3′; P2 = 3′-AAAAGCCATCAGTTGGTTGG-5′; P3 = 5′-AGCAACTGGAAGAGCTGGAA-3′; P4 = 3′-TGACAGGGTCCATGCTGTAG-5′.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Pax3 isoforms in sensory neurogenesis: expression and function in the ophthalmic trigeminal placode
    Article Snippet: Paragraph title: RNA isolation and RT-PCR ... Reverse transcription of RNA was performed using a Superscript Double-Stranded cDNA Synthesis Kit from Invitrogen with oligo primers.

    Sonication:

    Article Title: Ab initio construction of a eukaryotic transcriptome by massively parallel mRNA sequencing
    Article Snippet: The cDNA was synthesized by using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen) with SuperScript III (Invitrogen), 15-ng random hexamers (Invitrogen), and 20 units SUPERase·In (Ambion). .. Primer annealing was done at room temperature for 10 min followed by 1 h at 55 °C for first strand synthesis and 2 h at 16 °C for second strand synthesis; cDNA was sheared by sonication with 12 alternating cycles between “high intensity” (30 s; duty cycle, 20%; intensity, 10%; cycles per burst, 200) and “low intensity” (4 s; duty cycle, 5%; intensity, 10%; cycles per burst, 200) in the Frequency Sweeping mode (Covaris S2 machine).

    RNA Sequencing Assay:

    Article Title: Leptin Stimulates Cellular Glycolysis Through a STAT3 Dependent Mechanism in Tilapia
    Article Snippet: Paragraph title: RNA-seq and bioinformatic analyses of the pituitary transcriptome ... The cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen) and tagged with a 5′, four-nucleotide barcode.

    Article Title: Rsx, a metatherian RNA with Xist-like properties
    Article Snippet: Paragraph title: RNA-Seq ... Reaction mixture was purified using QIAquick PCR purification kit (Qiagen) and second strand synthesis was performed using SuperScript double-stranded cDNA synthesis kit (Invitrogen) as recommended by the manufacturer with the exception that 200 µM dTTP was replaced with 400 µM dUTP.

    Magnetic Beads:

    Article Title: Single-molecule long-read sequencing facilitates shrimp transcriptome research
    Article Snippet: Briefly, poly-(A) mRNA was isolated from total RNA using oligo (dT) magnetic beads and then fragmented into 200–700 bp pieces with fragmentation buffer. .. Double-stranded cDNAs were synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, USA) with random hexamer primers (Illumina, USA), following the manufacturer’s instructions.

    Isolation:

    Article Title: Leptin Stimulates Cellular Glycolysis Through a STAT3 Dependent Mechanism in Tilapia
    Article Snippet: Tissues from each well were pooled in 2 mL of RNAlater at 4°C until RNA isolation and Illumina cDNA library preparation. .. The cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen) and tagged with a 5′, four-nucleotide barcode.

    Article Title: Ab initio construction of a eukaryotic transcriptome by massively parallel mRNA sequencing
    Article Snippet: Total RNA and polyA+ RNA were isolated by using the RNeasy Midi Kit (Qiagen) and Poly(A) Purist kit (Ambion), respectively. .. The cDNA was synthesized by using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen) with SuperScript III (Invitrogen), 15-ng random hexamers (Invitrogen), and 20 units SUPERase·In (Ambion).

    Article Title: Single-molecule long-read sequencing facilitates shrimp transcriptome research
    Article Snippet: Briefly, poly-(A) mRNA was isolated from total RNA using oligo (dT) magnetic beads and then fragmented into 200–700 bp pieces with fragmentation buffer. .. Double-stranded cDNAs were synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, USA) with random hexamer primers (Illumina, USA), following the manufacturer’s instructions.

    Article Title: De novo assembly and characterization of root transcriptome using Illumina paired-end sequencing and development of cSSR markers in sweetpotato (Ipomoea batatas)
    Article Snippet: Briefly, poly (A) RNA was isolated from 20 μg of total RNA using Sera-mag Magnetic Oligo (dT) Beads (Illumina). .. Then the double-stranded cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen, Camarillo, CA) with random hexamer (N6) primers (Illumina).

    Article Title: Transcriptional Dysregulation in NIPBL and Cohesin Mutant Human Cells
    Article Snippet: .. RNA Isolation and Affymatrix Expression Array Hybridization Total RNA from each sample was extracted with the RNeasy Mini-kit (Qiagen), synthesis of double-stranded cDNA was performed using SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen), and cleaned up with GeneChip Sample Cleanup module (Affymetrix). .. The resulting products were then used to synthesize biotin-labeled cRNA with Enzo Bioarray High Yield RNA Transcript Labeling kit (Enzo Life Sciences) and further fragmented to 35–200-bp oligos.

    Article Title: Pax3 isoforms in sensory neurogenesis: expression and function in the ophthalmic trigeminal placode
    Article Snippet: Paragraph title: RNA isolation and RT-PCR ... Reverse transcription of RNA was performed using a Superscript Double-Stranded cDNA Synthesis Kit from Invitrogen with oligo primers.

    Size-exclusion Chromatography:

    Article Title: Rsx, a metatherian RNA with Xist-like properties
    Article Snippet: Reaction mixture was purified using QIAquick PCR purification kit (Qiagen) and second strand synthesis was performed using SuperScript double-stranded cDNA synthesis kit (Invitrogen) as recommended by the manufacturer with the exception that 200 µM dTTP was replaced with 400 µM dUTP. .. Double-stranded cDNA was fragmented using Bioruptor (Diagenode) (15 min, Low power, 30 sec ON, 30 sec OFF) in 50 µl volume.

    Labeling:

    Article Title: Comparative Transcriptional Profiling of Two Contrasting Barley Genotypes under Salinity Stress during the Seedling Stage
    Article Snippet: The SuperScript double-stranded cDNA synthesis kit (Invitrogen) was used to synthesize complementary DNA. .. A part of the double-stranded cDNA was used as a template to produce biotin-tagged cRNA by an Affymetrix GeneChip IVT labeling kit (Affymetrix).

    Article Title: Identification and functional clustering of global gene expression differences between human age-related cataract and clear lenses
    Article Snippet: .. First and second strand cDNAs were synthesized from 2–5 μg of total RNA using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen, Gaithersburg, MD) and the oligo-dT24 -T7 primer (5′-GGC CAG TGA ATT GTA ATA CGA CTC ACT AT-AGG GAG GCG G-3′) according to the manufacturer's instructions. cRNA was synthesized and labeled with biotinylated UTP and CTP by in vitro transcription using the T7 promoter coupled double-stranded cDNA as a template and the T7 RNA Transcript Labeling Kit (ENZO Diagnostics Inc., Farmingdale, NY). .. Briefly, double-stranded cDNAs synthesized from the previous steps were washed twice with 70% ethanol and resuspended in 22 μl of RNase-free H2 O.

    Article Title: Transcriptional Profiling of Lipopolysaccharide-Induced Acute Lung Injury
    Article Snippet: Single-stranded and then double-stranded cDNAs were synthesized from the poly(A+ ) mRNA present in the purified total RNA by use of a SuperScript double-stranded cDNA synthesis kit (Invitrogen Corp.) and poly(T) nucleotide primers that contained a sequence recognized by T7 RNA polymerase. .. The resulting double-stranded cDNAs were used as templates to generate biotinylated cRNAs by in vitro transcription reactions with a Bio-Array High-Yield RNA transcript labeling kit (Enzo Diagnostics Inc., Farmingdale, N.Y.).

    Article Title: Transcriptional Dysregulation in NIPBL and Cohesin Mutant Human Cells
    Article Snippet: RNA Isolation and Affymatrix Expression Array Hybridization Total RNA from each sample was extracted with the RNeasy Mini-kit (Qiagen), synthesis of double-stranded cDNA was performed using SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen), and cleaned up with GeneChip Sample Cleanup module (Affymetrix). .. The resulting products were then used to synthesize biotin-labeled cRNA with Enzo Bioarray High Yield RNA Transcript Labeling kit (Enzo Life Sciences) and further fragmented to 35–200-bp oligos.

    Purification:

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: .. Amplified RNA was converted to double-stranded cDNA using the SuperScript III First-Strand Synthesis System (Life Technologies) with priming via random hexamers for the first-strand synthesis and the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for the second-strand synthesis. cDNA in the 0.3–3.0-kb size range was purified from agarose gels using the QIAquick Gel Extraction Kit reagents and protocols (Qiagen) and digested with BpmI for 2–3 h at 37 °C to remove poly(A) tails. .. Following BpmI digestion, cDNA was purified with Ampure XP beads (Beckman Coulter, Brea, CA, USA) and used directly for pyrosequencing.

    Article Title: Transcriptional Profiling of Lipopolysaccharide-Induced Acute Lung Injury
    Article Snippet: .. Single-stranded and then double-stranded cDNAs were synthesized from the poly(A+ ) mRNA present in the purified total RNA by use of a SuperScript double-stranded cDNA synthesis kit (Invitrogen Corp.) and poly(T) nucleotide primers that contained a sequence recognized by T7 RNA polymerase. .. The resulting double-stranded cDNAs were used as templates to generate biotinylated cRNAs by in vitro transcription reactions with a Bio-Array High-Yield RNA transcript labeling kit (Enzo Diagnostics Inc., Farmingdale, N.Y.).

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: RNA was purified from filters using the Trizol reagent (Life Technologies), treated with DNase (Qiagen) and cleaned with the RNeasy MinElute Kit (Qiagen). .. OligoDT primers were used for cDNA synthesis (SuperScript III, Life Technologies) followed by the the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for second-strand synthesis.

    Article Title: Rsx, a metatherian RNA with Xist-like properties
    Article Snippet: .. Reaction mixture was purified using QIAquick PCR purification kit (Qiagen) and second strand synthesis was performed using SuperScript double-stranded cDNA synthesis kit (Invitrogen) as recommended by the manufacturer with the exception that 200 µM dTTP was replaced with 400 µM dUTP. .. Double-stranded cDNA was fragmented using Bioruptor (Diagenode) (15 min, Low power, 30 sec ON, 30 sec OFF) in 50 µl volume.

    Article Title: De novo assembly and characterization of root transcriptome using Illumina paired-end sequencing and development of cSSR markers in sweetpotato (Ipomoea batatas)
    Article Snippet: To avoid priming bias when synthesizing cDNA, the purified mRNA was first fragmented into small pieces (100-400 bp) using divalent cations at 94°C for exactly 5 minutes. .. Then the double-stranded cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen, Camarillo, CA) with random hexamer (N6) primers (Illumina).

    Article Title: Experimental Incubations Elicit Profound Changes in Community Transcription in OMZ Bacterioplankton
    Article Snippet: .. Antisense RNA (∼5–10 µg aliquot) was then converted to double-stranded cDNA using the SuperScript® III First-Strand Synthesis System (Invitrogen) with priming via random hexamers for first-strand synthesis, and the SuperScript™ Double-Stranded cDNA synthesis kit (Invitrogen) for second-strand synthesis. cDNA was then purified with the QIAquick PCR purification kit (Qiagen), restriction-digested with BpmI to remove poly(A) tails, and used directly for pyrosequencing .. Pyrosequencing Using standard protocols (454 Life Sciences, Roche), poly(A)-removed cDNA was purified via the AMPure® kit (Agencourt®) and used to generate single-stranded DNA libraries for use as template in emulsion PCR.

    Article Title: Pax3 isoforms in sensory neurogenesis: expression and function in the ophthalmic trigeminal placode
    Article Snippet: Total RNA was isolated from all tissue samples using 2-mercaptoethanol and purified using the PureLink Micro-to-Midi Total RNA Purification System kit (Invitrogen). .. Reverse transcription of RNA was performed using a Superscript Double-Stranded cDNA Synthesis Kit from Invitrogen with oligo primers.

    Sequencing:

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: Briefly, amplified rRNA-subtracted RNA was converted to double-stranded cDNA via reverse transcription primed with a modified Oligo(dT) primer containing a promoter sequence for T7 RNA polymerase and a recognition site for the restriction enzyme BpmI ( ). cDNA was then subjected to two rounds of in vitro transcription at 37 °C for 14 h yielding large quantities (100–200 μg) of single-stranded antisense RNA. .. Amplified RNA was converted to double-stranded cDNA using the SuperScript III First-Strand Synthesis System (Life Technologies) with priming via random hexamers for the first-strand synthesis and the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for the second-strand synthesis. cDNA in the 0.3–3.0-kb size range was purified from agarose gels using the QIAquick Gel Extraction Kit reagents and protocols (Qiagen) and digested with BpmI for 2–3 h at 37 °C to remove poly(A) tails.

    Article Title: Leptin Stimulates Cellular Glycolysis Through a STAT3 Dependent Mechanism in Tilapia
    Article Snippet: The cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen) and tagged with a 5′, four-nucleotide barcode. .. Sequencing was performed in triplicate at the Hawaii Institute of Marine Biology Genetics core facility on an Illumina MiSeq platform, using 150 bp, paired-end reads.

    Article Title: Transcriptional Profiling of Lipopolysaccharide-Induced Acute Lung Injury
    Article Snippet: .. Single-stranded and then double-stranded cDNAs were synthesized from the poly(A+ ) mRNA present in the purified total RNA by use of a SuperScript double-stranded cDNA synthesis kit (Invitrogen Corp.) and poly(T) nucleotide primers that contained a sequence recognized by T7 RNA polymerase. .. The resulting double-stranded cDNAs were used as templates to generate biotinylated cRNAs by in vitro transcription reactions with a Bio-Array High-Yield RNA transcript labeling kit (Enzo Diagnostics Inc., Farmingdale, N.Y.).

    Article Title: Ab initio construction of a eukaryotic transcriptome by massively parallel mRNA sequencing
    Article Snippet: The cDNA was synthesized by using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen) with SuperScript III (Invitrogen), 15-ng random hexamers (Invitrogen), and 20 units SUPERase·In (Ambion). .. Adapters for Illumina sequencing were added following the instructions provided, except that 5 times less adapter mix was ligated to the cDNAs and PCR primers were removed by digestion with RecJ (New England Biolabs).

    Article Title: Single-molecule long-read sequencing facilitates shrimp transcriptome research
    Article Snippet: Paragraph title: Illumina library construction and sequencing ... Double-stranded cDNAs were synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, USA) with random hexamer primers (Illumina, USA), following the manufacturer’s instructions.

    Article Title: Rsx, a metatherian RNA with Xist-like properties
    Article Snippet: Reaction mixture was purified using QIAquick PCR purification kit (Qiagen) and second strand synthesis was performed using SuperScript double-stranded cDNA synthesis kit (Invitrogen) as recommended by the manufacturer with the exception that 200 µM dTTP was replaced with 400 µM dUTP. .. Sequencing libraries were prepared from fragmented cDNA using NEBNext DNA sample preparation kit (New England Biolabs) and Illumina PE adapters (PE-102–1003).

    Article Title: De novo assembly and characterization of root transcriptome using Illumina paired-end sequencing and development of cSSR markers in sweetpotato (Ipomoea batatas)
    Article Snippet: Paragraph title: cDNA library construction and sequencing ... Then the double-stranded cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen, Camarillo, CA) with random hexamer (N6) primers (Illumina).

    Article Title: Experimental Incubations Elicit Profound Changes in Community Transcription in OMZ Bacterioplankton
    Article Snippet: Briefly, total RNA was polyadenylated using Escherichia coli poly(A) polymerase and converted to double-stranded cDNA via reverse transcription primed with an oligo(dT) primer containing a promoter sequence for T7 RNA polymerase and a recognition site for the restriction enzyme BpmI. cDNA was transcribed in vitro (37°C, 12–14 hr) to produce microgram quantities of single-stranded antisense RNA. .. Antisense RNA (∼5–10 µg aliquot) was then converted to double-stranded cDNA using the SuperScript® III First-Strand Synthesis System (Invitrogen) with priming via random hexamers for first-strand synthesis, and the SuperScript™ Double-Stranded cDNA synthesis kit (Invitrogen) for second-strand synthesis. cDNA was then purified with the QIAquick PCR purification kit (Qiagen), restriction-digested with BpmI to remove poly(A) tails, and used directly for pyrosequencing

    cDNA Library Assay:

    Article Title: Leptin Stimulates Cellular Glycolysis Through a STAT3 Dependent Mechanism in Tilapia
    Article Snippet: Pituitary mRNA (10 μg) was submitted to the North Carolina State University Genomic Sciences Laboratory (Raleigh, NC) for Illumina cDNA library construction. .. The cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen) and tagged with a 5′, four-nucleotide barcode.

    Article Title: De novo assembly and characterization of root transcriptome using Illumina paired-end sequencing and development of cSSR markers in sweetpotato (Ipomoea batatas)
    Article Snippet: Paragraph title: cDNA library construction and sequencing ... Then the double-stranded cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen, Camarillo, CA) with random hexamer (N6) primers (Illumina).

    Activated Clotting Time Assay:

    Article Title: Identification and functional clustering of global gene expression differences between human age-related cataract and clear lenses
    Article Snippet: .. First and second strand cDNAs were synthesized from 2–5 μg of total RNA using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen, Gaithersburg, MD) and the oligo-dT24 -T7 primer (5′-GGC CAG TGA ATT GTA ATA CGA CTC ACT AT-AGG GAG GCG G-3′) according to the manufacturer's instructions. cRNA was synthesized and labeled with biotinylated UTP and CTP by in vitro transcription using the T7 promoter coupled double-stranded cDNA as a template and the T7 RNA Transcript Labeling Kit (ENZO Diagnostics Inc., Farmingdale, NY). .. Briefly, double-stranded cDNAs synthesized from the previous steps were washed twice with 70% ethanol and resuspended in 22 μl of RNase-free H2 O.

    Chromatin Immunoprecipitation:

    Article Title: Comparative Transcriptional Profiling of Two Contrasting Barley Genotypes under Salinity Stress during the Seedling Stage
    Article Snippet: Paragraph title: 2.3. RNA Preparations and Gene Chip Hybridization ... The SuperScript double-stranded cDNA synthesis kit (Invitrogen) was used to synthesize complementary DNA.

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: OligoDT primers were used for cDNA synthesis (SuperScript III, Life Technologies) followed by the the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for second-strand synthesis. .. A high-sensitivity DNA Assay chip was used to assess quality (Agilent; Santa Clara, CA, USA).

    RNA Extraction:

    Article Title: Ab initio construction of a eukaryotic transcriptome by massively parallel mRNA sequencing
    Article Snippet: Paragraph title: RNA extraction and library preparation. ... The cDNA was synthesized by using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen) with SuperScript III (Invitrogen), 15-ng random hexamers (Invitrogen), and 20 units SUPERase·In (Ambion).

    Sample Prep:

    Article Title: Single-molecule long-read sequencing facilitates shrimp transcriptome research
    Article Snippet: Illumina library construction and sequencing The Illumina libraries used to correct the FLNC reads were constructed with the Tru-Seq RNA sample Prep kit (Illumina, USA). .. Double-stranded cDNAs were synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, USA) with random hexamer primers (Illumina, USA), following the manufacturer’s instructions.

    Article Title: Rsx, a metatherian RNA with Xist-like properties
    Article Snippet: Reaction mixture was purified using QIAquick PCR purification kit (Qiagen) and second strand synthesis was performed using SuperScript double-stranded cDNA synthesis kit (Invitrogen) as recommended by the manufacturer with the exception that 200 µM dTTP was replaced with 400 µM dUTP. .. Sequencing libraries were prepared from fragmented cDNA using NEBNext DNA sample preparation kit (New England Biolabs) and Illumina PE adapters (PE-102–1003).

    In Vitro:

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: Briefly, amplified rRNA-subtracted RNA was converted to double-stranded cDNA via reverse transcription primed with a modified Oligo(dT) primer containing a promoter sequence for T7 RNA polymerase and a recognition site for the restriction enzyme BpmI ( ). cDNA was then subjected to two rounds of in vitro transcription at 37 °C for 14 h yielding large quantities (100–200 μg) of single-stranded antisense RNA. .. Amplified RNA was converted to double-stranded cDNA using the SuperScript III First-Strand Synthesis System (Life Technologies) with priming via random hexamers for the first-strand synthesis and the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for the second-strand synthesis. cDNA in the 0.3–3.0-kb size range was purified from agarose gels using the QIAquick Gel Extraction Kit reagents and protocols (Qiagen) and digested with BpmI for 2–3 h at 37 °C to remove poly(A) tails.

    Article Title: Identification and functional clustering of global gene expression differences between human age-related cataract and clear lenses
    Article Snippet: .. First and second strand cDNAs were synthesized from 2–5 μg of total RNA using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen, Gaithersburg, MD) and the oligo-dT24 -T7 primer (5′-GGC CAG TGA ATT GTA ATA CGA CTC ACT AT-AGG GAG GCG G-3′) according to the manufacturer's instructions. cRNA was synthesized and labeled with biotinylated UTP and CTP by in vitro transcription using the T7 promoter coupled double-stranded cDNA as a template and the T7 RNA Transcript Labeling Kit (ENZO Diagnostics Inc., Farmingdale, NY). .. Briefly, double-stranded cDNAs synthesized from the previous steps were washed twice with 70% ethanol and resuspended in 22 μl of RNase-free H2 O.

    Article Title: Transcriptional Profiling of Lipopolysaccharide-Induced Acute Lung Injury
    Article Snippet: Single-stranded and then double-stranded cDNAs were synthesized from the poly(A+ ) mRNA present in the purified total RNA by use of a SuperScript double-stranded cDNA synthesis kit (Invitrogen Corp.) and poly(T) nucleotide primers that contained a sequence recognized by T7 RNA polymerase. .. The resulting double-stranded cDNAs were used as templates to generate biotinylated cRNAs by in vitro transcription reactions with a Bio-Array High-Yield RNA transcript labeling kit (Enzo Diagnostics Inc., Farmingdale, N.Y.).

    Article Title: Experimental Incubations Elicit Profound Changes in Community Transcription in OMZ Bacterioplankton
    Article Snippet: Briefly, total RNA was polyadenylated using Escherichia coli poly(A) polymerase and converted to double-stranded cDNA via reverse transcription primed with an oligo(dT) primer containing a promoter sequence for T7 RNA polymerase and a recognition site for the restriction enzyme BpmI. cDNA was transcribed in vitro (37°C, 12–14 hr) to produce microgram quantities of single-stranded antisense RNA. .. Antisense RNA (∼5–10 µg aliquot) was then converted to double-stranded cDNA using the SuperScript® III First-Strand Synthesis System (Invitrogen) with priming via random hexamers for first-strand synthesis, and the SuperScript™ Double-Stranded cDNA synthesis kit (Invitrogen) for second-strand synthesis. cDNA was then purified with the QIAquick PCR purification kit (Qiagen), restriction-digested with BpmI to remove poly(A) tails, and used directly for pyrosequencing

    Random Hexamer Labeling:

    Article Title: Single-molecule long-read sequencing facilitates shrimp transcriptome research
    Article Snippet: .. Double-stranded cDNAs were synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, USA) with random hexamer primers (Illumina, USA), following the manufacturer’s instructions. .. Synthesized cDNAs were gen-purified and amplified with PCR.

    Article Title: De novo assembly and characterization of root transcriptome using Illumina paired-end sequencing and development of cSSR markers in sweetpotato (Ipomoea batatas)
    Article Snippet: .. Then the double-stranded cDNA was synthesized using the SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen, Camarillo, CA) with random hexamer (N6) primers (Illumina). .. The synthesized cDNA was subjected to end-repair and phosphorylation using T4 DNA polymerase, Klenow DNA polymerase and T4 PNK.

    Spectrophotometry:

    Article Title: Comparative Transcriptional Profiling of Two Contrasting Barley Genotypes under Salinity Stress during the Seedling Stage
    Article Snippet: The quality of RNA was analyzed on a denaturing formaldehyde gel and confirmed by measuring the ratio of A260/A280 with the NanoDropND-1000 spectrophotometer. .. The SuperScript double-stranded cDNA synthesis kit (Invitrogen) was used to synthesize complementary DNA.

    Sampling:

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: Paragraph title: Oceanographic sampling ... Amplified RNA was converted to double-stranded cDNA using the SuperScript III First-Strand Synthesis System (Life Technologies) with priming via random hexamers for the first-strand synthesis and the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for the second-strand synthesis. cDNA in the 0.3–3.0-kb size range was purified from agarose gels using the QIAquick Gel Extraction Kit reagents and protocols (Qiagen) and digested with BpmI for 2–3 h at 37 °C to remove poly(A) tails.

    Concentration Assay:

    Article Title: Transcriptional Dysregulation in NIPBL and Cohesin Mutant Human Cells
    Article Snippet: RNA Isolation and Affymatrix Expression Array Hybridization Total RNA from each sample was extracted with the RNeasy Mini-kit (Qiagen), synthesis of double-stranded cDNA was performed using SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen), and cleaned up with GeneChip Sample Cleanup module (Affymetrix). .. 30 µl fragmented cRNA at the concentration of 500 ng/µl was sent for hybridization in the microarray facility at The Children's Hospital of Philadelphia.

    High Throughput Screening Assay:

    Article Title: Single-molecule long-read sequencing facilitates shrimp transcriptome research
    Article Snippet: Double-stranded cDNAs were synthesized using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, USA) with random hexamer primers (Illumina, USA), following the manufacturer’s instructions. .. 2500 high-throughput sequencer.

    Gel Extraction:

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: .. Amplified RNA was converted to double-stranded cDNA using the SuperScript III First-Strand Synthesis System (Life Technologies) with priming via random hexamers for the first-strand synthesis and the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for the second-strand synthesis. cDNA in the 0.3–3.0-kb size range was purified from agarose gels using the QIAquick Gel Extraction Kit reagents and protocols (Qiagen) and digested with BpmI for 2–3 h at 37 °C to remove poly(A) tails. .. Following BpmI digestion, cDNA was purified with Ampure XP beads (Beckman Coulter, Brea, CA, USA) and used directly for pyrosequencing.

    Article Title: Genomes and gene expression across light and productivity gradients in eastern subtropical Pacific microbial communities
    Article Snippet: OligoDT primers were used for cDNA synthesis (SuperScript III, Life Technologies) followed by the the SuperScript Double-Stranded cDNA synthesis kit (Life Technologies) for second-strand synthesis. .. Size-selected cDNA (0.5–1 kb) was purified using the QiaQuick gel extraction kit (Qiagen) and further purified with AMPure (Beckman Coulter).

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    Thermo Fisher scientific simple • taqman universal master mix ii
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    Thermo Fisher superscript ii first strand cdna synthesis kit
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