protoscript ii reverse transcriptase  (New England Biolabs)


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    Name:
    ProtoScript II Reverse Transcriptase
    Description:
    ProtoScript II Reverse Transcriptase 10 000 units
    Catalog Number:
    m0368l
    Price:
    180
    Size:
    10 000 units
    Category:
    Reverse Transcriptases
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    New England Biolabs protoscript ii reverse transcriptase
    ProtoScript II Reverse Transcriptase
    ProtoScript II Reverse Transcriptase 10 000 units
    https://www.bioz.com/result/protoscript ii reverse transcriptase/product/New England Biolabs
    Average 95 stars, based on 113 article reviews
    Price from $9.99 to $1999.99
    protoscript ii reverse transcriptase - by Bioz Stars, 2020-02
    95/100 stars

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    Amplification:

    Article Title: A novel hybrid SCCmec-mecC region in Staphylococcus sciuri
    Article Snippet: After an additional DNAse step using RQ1 RNase-Free DNase (Promega, UK), cDNA was synthesized using ProtoScript® II Reverse Transcriptase (NEB, UK) and a Random Hexamer primer (Fisher Scientific, UK) following the manufacturer's standard protocol. .. Controls without reverse transcriptase were generated for all samples and showed no amplification in the subsequent PCRs. cDNA was used undiluted in a standard PCR for the detection of mecC (mecC-Uni-F/R), mecA (MecA1/A2) and mecA1 (mecA1-spec-F/R) (Table ).

    Article Title: Global Transcriptome Analysis of Aedes aegypti Mosquitoes in Response to Zika Virus Infection
    Article Snippet: Fragmented poly(A)+ RNA samples were converted to cDNA by random primed synthesis using ProtoScript II reverse transcriptase (New England Biolabs). .. After ligation, the template DNAs were amplified (5 to 9 cycles) using primers specific to each of the noncomplementary sequences in the adapters.

    Positive Control:

    Article Title: A novel hybrid SCCmec-mecC region in Staphylococcus sciuri
    Article Snippet: After an additional DNAse step using RQ1 RNase-Free DNase (Promega, UK), cDNA was synthesized using ProtoScript® II Reverse Transcriptase (NEB, UK) and a Random Hexamer primer (Fisher Scientific, UK) following the manufacturer's standard protocol. .. A PCR for 16S rRNA (Uni-16s-Ctrl-F/R) was also carried out as a positive control for cDNA synthesis (Table ).

    Synthesized:

    Article Title: Overexpression of TET dioxygenases in seminomas associates with low levels of DNA methylation and hydroxymethylation
    Article Snippet: .. Alternatively, total RNA was isolated using RNA MicroPrep™ (Zymo Research) according to manufacturer’s instructions. cDNA for the quantitative RT-PCR (qRT-PCR) analysis was synthesized using Protoscript II reverse transcriptase (NEB) and random hexanucleotides (Promega). ..

    Article Title: A novel hybrid SCCmec-mecC region in Staphylococcus sciuri
    Article Snippet: .. After an additional DNAse step using RQ1 RNase-Free DNase (Promega, UK), cDNA was synthesized using ProtoScript® II Reverse Transcriptase (NEB, UK) and a Random Hexamer primer (Fisher Scientific, UK) following the manufacturer's standard protocol. .. Controls without reverse transcriptase were generated for all samples and showed no amplification in the subsequent PCRs. cDNA was used undiluted in a standard PCR for the detection of mecC (mecC-Uni-F/R), mecA (MecA1/A2) and mecA1 (mecA1-spec-F/R) (Table ).

    Article Title: Heterogeneity of tumor-infiltrating lymphocytes ascribed to local immune status rather than neoantigens by multi-omics analysis of glioblastoma multiforme
    Article Snippet: .. TCRB-specific cDNA was synthesized from 0.5 μg of total RNA using ProtoScript II Reverse Transcriptase (NEB, UK) and primer TRBCRo. .. For the first PCR step, cDNA templates were pre-amplified using a multiplex PCR 5x Master Mix (NEB, UK) with multiple Vβ region primers (TRBV1Fo-TRBV30Fo) and a Cβ region primer (TRBCRo primer).

    Article Title: Streptococcus suis contains multiple phase-variable methyltransferases that show a discrete lineage distribution
    Article Snippet: .. Prior to reverse transcription (RT), RNA was treated with heat-labile dsDNAse (Thermo Fisher catalogue number EN0771) according to manufacturer’s instructions, and used directly in the RT reaction. cDNA was synthesized from DNAse-treated RNA using Protoscript II Reverse Transcriptase (New England Biolabs) according to manufacturer’s instructions. ..

    Article Title: Coordination of RNA Polymerase II Pausing and 3′ End Processing Factor Recruitment with Alternative Polyadenylation
    Article Snippet: .. Total RNA was extracted by use of TRIzol. cDNA was synthesized using Protoscript II reverse transcriptase (NEB) with the dT18-XbaKpnBam primer for 3′ rapid amplification of cDNA ends (RACE) ( , ). .. 3′ RACE was carried out using nested forward primers and the XbaKpnBam reverse primer, as previously described ( , ).

    Quantitative RT-PCR:

    Article Title: In Vitro Study on Anti-Hepatitis C Virus Activity of Spatholobus suberectus Dunn
    Article Snippet: Paragraph title: 4.9. Extraction of Total RNA and Quantitative Real-Time-PCR (qRT-PCR) Analysis ... One microgram of total RNA was reverse transcribed into cDNA with ProtoScript® II Reverse Transcriptase (New England Biolabs, Ipswich, MA, USA). qPCR was performed in triplicates for all samples using the Mx 3005P qPCR system (Agilent, Santa Clara, CA, USA) with SYBR Master Mix (Bio-Rad, Hercules, CA, USA).

    Article Title: Overexpression of TET dioxygenases in seminomas associates with low levels of DNA methylation and hydroxymethylation
    Article Snippet: .. Alternatively, total RNA was isolated using RNA MicroPrep™ (Zymo Research) according to manufacturer’s instructions. cDNA for the quantitative RT-PCR (qRT-PCR) analysis was synthesized using Protoscript II reverse transcriptase (NEB) and random hexanucleotides (Promega). ..

    Article Title: HoxBlinc RNA recruits Set1/MLL complexes to activate Hox gene expression patterns and mesoderm lineage development
    Article Snippet: .. 2 µg of RNA was reverse transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs). cDNA was analyzed by real-time PCR (qRT-PCR) using the SYBR Green PCR master mix and the Bio-Rad CFX96 system (Bio-Rad). ..

    SYBR Green Assay:

    Article Title: Ectopic expression of a cyanobacterial flavodoxin in creeping bentgrass impacts plant development and confers broad abiotic stress tolerance
    Article Snippet: RNA isolation, cDNA synthesis, qPCR, RT‐PCR and northern blot One hundred milligrams of young leaf tissues was employed for total RNA isolation using Trizol reagent (Invitrogen, Carlsbad, CA), and 2 μg of total RNAs was reverse‐transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs, GA) following manufacturer's instructions. .. RT‐PCR for FRN‐Fld expression determination was conducted using the first‐strand cDNAs and gene‐specific primers (Table S1). qPCR was carried out on an iCycler iQ system (Bio‐Rad, Hercules, CA) in 25 μL of PCR solution containing 10 ppm SYBR Green I, 200 μm dNTPs, 1× PCR buffer, 1.5 mm MgCl2 , Taq DNA polymerase and 40 nM each primer.

    Article Title: HoxBlinc RNA recruits Set1/MLL complexes to activate Hox gene expression patterns and mesoderm lineage development
    Article Snippet: .. 2 µg of RNA was reverse transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs). cDNA was analyzed by real-time PCR (qRT-PCR) using the SYBR Green PCR master mix and the Bio-Rad CFX96 system (Bio-Rad). ..

    Article Title: Design, synthesis, and biological evaluation of inhibitors of the NADPH oxidase, Nox4
    Article Snippet: Reactions were carried out at 42 °C, using random 15-mer primers and ProtoScript II reverse transcriptase (New England Biolabs, Beverly, MA). .. Quantitative PCR was performed with a LightCycler instrument in glass capillaries (Roche Applied Science, Indianapolis, IN), using Platinum Taq DNA polymerase and SYBR green (Invitrogen, Carlsbad, CA).

    Incubation:

    Article Title: 3′ end additions by T7 RNA polymerase are RNA self-templated, distributive and diverse in character—RNA-Seq analyses
    Article Snippet: .. The reverse transcription reactions were performed in a 50 μl reaction volume containing 0.4 μM reverse transcriptase primer, 0.5 mM each deoxynucleoside triphosphate (dNTP), 10 mM dithiothreitol (DTT) and 2 μl ProtoScript® II Reverse Transcriptase (New England BioLabs) at 200 U μl−1 by incubation at 42°C for 1 h. .. Amplification and quantification of the library cDNA for each experiment was amplified by Phusion® High-Fidelity DNA Polymerase (New England BioLabs) using primers based on Illumina primers for TruSeq Small RNA Library, with a final concentration of 0.4 μM (primers were purchased from IDT and are shown in ).

    Article Title: Superinfection with Woodchuck Hepatitis Virus Strain WHVNY of Livers Chronically Infected with Strain WHV7
    Article Snippet: An additional 2 U of Turbo DNase then was added to each sample, and the samples were incubated at 37°C for another hour. .. The cDNA synthesis reaction was started by adding 200 U of ProtoScript II reverse transcriptase (RT; New England BioLabs [NEB]), 2 μl of 10× ProtoScript II buffer, and dithiothreitol (DTT) (to a final concentration of 10 mM).

    Expressing:

    Article Title: Aquaporin 5 Expression in Mouse Mammary Gland Cells Is Not Driven by Promoter Methylation
    Article Snippet: Paragraph title: 2.9. AQP5 Expression in Human Mammary Gland MCF-7 Cells ... The pellet was washed two times with PBS before mRNA extraction with the Quick-RNA MiniPrep Kit (Zymo Research); cDNA synthesis was performed with the ProtoScript II Reverse Transcriptase (NEB), both according to manufacturer's instructions.

    Article Title: Ectopic expression of a cyanobacterial flavodoxin in creeping bentgrass impacts plant development and confers broad abiotic stress tolerance
    Article Snippet: RNA isolation, cDNA synthesis, qPCR, RT‐PCR and northern blot One hundred milligrams of young leaf tissues was employed for total RNA isolation using Trizol reagent (Invitrogen, Carlsbad, CA), and 2 μg of total RNAs was reverse‐transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs, GA) following manufacturer's instructions. .. RT‐PCR for FRN‐Fld expression determination was conducted using the first‐strand cDNAs and gene‐specific primers (Table S1). qPCR was carried out on an iCycler iQ system (Bio‐Rad, Hercules, CA) in 25 μL of PCR solution containing 10 ppm SYBR Green I, 200 μm dNTPs, 1× PCR buffer, 1.5 mm MgCl2 , Taq DNA polymerase and 40 nM each primer.

    Article Title: A novel hybrid SCCmec-mecC region in Staphylococcus sciuri
    Article Snippet: Paragraph title: Transcriptional analysis of mec gene expression by RT–PCR ... After an additional DNAse step using RQ1 RNase-Free DNase (Promega, UK), cDNA was synthesized using ProtoScript® II Reverse Transcriptase (NEB, UK) and a Random Hexamer primer (Fisher Scientific, UK) following the manufacturer's standard protocol.

    Modification:

    Article Title: Heterogeneity of tumor-infiltrating lymphocytes ascribed to local immune status rather than neoantigens by multi-omics analysis of glioblastoma multiforme
    Article Snippet: TCR sequencing The TCR sequences were obtained via a series of three nested PCR reactions using the primers as described in ref. (Table ) with a slightly modified protocol of the ARM-PCR procedure , . .. TCRB-specific cDNA was synthesized from 0.5 μg of total RNA using ProtoScript II Reverse Transcriptase (NEB, UK) and primer TRBCRo.

    Rapid Amplification of cDNA Ends:

    Article Title: Coordination of RNA Polymerase II Pausing and 3′ End Processing Factor Recruitment with Alternative Polyadenylation
    Article Snippet: .. Total RNA was extracted by use of TRIzol. cDNA was synthesized using Protoscript II reverse transcriptase (NEB) with the dT18-XbaKpnBam primer for 3′ rapid amplification of cDNA ends (RACE) ( , ). .. 3′ RACE was carried out using nested forward primers and the XbaKpnBam reverse primer, as previously described ( , ).

    Ligation:

    Article Title: Global Transcriptome Analysis of Aedes aegypti Mosquitoes in Response to Zika Virus Infection
    Article Snippet: Fragmented poly(A)+ RNA samples were converted to cDNA by random primed synthesis using ProtoScript II reverse transcriptase (New England Biolabs). .. After ligation, the template DNAs were amplified (5 to 9 cycles) using primers specific to each of the noncomplementary sequences in the adapters.

    Northern Blot:

    Article Title: Ectopic expression of a cyanobacterial flavodoxin in creeping bentgrass impacts plant development and confers broad abiotic stress tolerance
    Article Snippet: .. RNA isolation, cDNA synthesis, qPCR, RT‐PCR and northern blot One hundred milligrams of young leaf tissues was employed for total RNA isolation using Trizol reagent (Invitrogen, Carlsbad, CA), and 2 μg of total RNAs was reverse‐transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs, GA) following manufacturer's instructions. ..

    Article Title: HoxBlinc RNA recruits Set1/MLL complexes to activate Hox gene expression patterns and mesoderm lineage development
    Article Snippet: Paragraph title: Northern blot and RT-qPCR ... 2 µg of RNA was reverse transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs). cDNA was analyzed by real-time PCR (qRT-PCR) using the SYBR Green PCR master mix and the Bio-Rad CFX96 system (Bio-Rad).

    Cell Culture:

    Article Title: Aquaporin 5 Expression in Mouse Mammary Gland Cells Is Not Driven by Promoter Methylation
    Article Snippet: Cells were seeded on 24 well plates and cultured for 2 days (yielding subconfluent cell layers with approximately 50% confluence) or 5 days (yielding confluent cell layers). .. The pellet was washed two times with PBS before mRNA extraction with the Quick-RNA MiniPrep Kit (Zymo Research); cDNA synthesis was performed with the ProtoScript II Reverse Transcriptase (NEB), both according to manufacturer's instructions.

    Article Title: Design, synthesis, and biological evaluation of inhibitors of the NADPH oxidase, Nox4
    Article Snippet: Total RNA samples (2 μg), purified from cultured MASMs, using the RNeasy Plus kit (Qiagen, Chatsworth, CA), were used to prepare cDNA. .. Reactions were carried out at 42 °C, using random 15-mer primers and ProtoScript II reverse transcriptase (New England Biolabs, Beverly, MA).

    Generated:

    Article Title: In Vitro Study on Anti-Hepatitis C Virus Activity of Spatholobus suberectus Dunn
    Article Snippet: One microgram of total RNA was reverse transcribed into cDNA with ProtoScript® II Reverse Transcriptase (New England Biolabs, Ipswich, MA, USA). qPCR was performed in triplicates for all samples using the Mx 3005P qPCR system (Agilent, Santa Clara, CA, USA) with SYBR Master Mix (Bio-Rad, Hercules, CA, USA). .. Absolute copy number of HCV RNA was determined with the standard curve generated by using the linearized pFK-I389/NS3-3′ plasmid bearing the HCV genome and 18S rRNA was included as a control.

    Article Title: A novel hybrid SCCmec-mecC region in Staphylococcus sciuri
    Article Snippet: After an additional DNAse step using RQ1 RNase-Free DNase (Promega, UK), cDNA was synthesized using ProtoScript® II Reverse Transcriptase (NEB, UK) and a Random Hexamer primer (Fisher Scientific, UK) following the manufacturer's standard protocol. .. Controls without reverse transcriptase were generated for all samples and showed no amplification in the subsequent PCRs. cDNA was used undiluted in a standard PCR for the detection of mecC (mecC-Uni-F/R), mecA (MecA1/A2) and mecA1 (mecA1-spec-F/R) (Table ).

    Article Title: Identification of radiation responsive genes and transcriptome profiling via complete RNA sequencing in a stable radioresistant U87 glioblastoma model
    Article Snippet: First strand complementary deoxyribonucleic acid (cDNA) syntheses were performed at 25° C for 10 minutes, 42° C for 15 minutes, and 70° C for 15 minutes, using random hexameres and ProtoScript II Reverse Transcriptase (New England BioLabs Inc., Ipswich, MA, USA). .. In a second strand cDNA synthesis, the RNA templates were removed and a second replacement strand was generated by incorporating deoxyuridine triphosphate (in place of deoxythymidine triphosphate, to keep strand information) to generate ds cDNA.

    Polymerase Chain Reaction:

    Article Title: Ectopic expression of a cyanobacterial flavodoxin in creeping bentgrass impacts plant development and confers broad abiotic stress tolerance
    Article Snippet: RNA isolation, cDNA synthesis, qPCR, RT‐PCR and northern blot One hundred milligrams of young leaf tissues was employed for total RNA isolation using Trizol reagent (Invitrogen, Carlsbad, CA), and 2 μg of total RNAs was reverse‐transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs, GA) following manufacturer's instructions. .. RT‐PCR for FRN‐Fld expression determination was conducted using the first‐strand cDNAs and gene‐specific primers (Table S1). qPCR was carried out on an iCycler iQ system (Bio‐Rad, Hercules, CA) in 25 μL of PCR solution containing 10 ppm SYBR Green I, 200 μm dNTPs, 1× PCR buffer, 1.5 mm MgCl2 , Taq DNA polymerase and 40 nM each primer.

    Article Title: A novel hybrid SCCmec-mecC region in Staphylococcus sciuri
    Article Snippet: After an additional DNAse step using RQ1 RNase-Free DNase (Promega, UK), cDNA was synthesized using ProtoScript® II Reverse Transcriptase (NEB, UK) and a Random Hexamer primer (Fisher Scientific, UK) following the manufacturer's standard protocol. .. Controls without reverse transcriptase were generated for all samples and showed no amplification in the subsequent PCRs. cDNA was used undiluted in a standard PCR for the detection of mecC (mecC-Uni-F/R), mecA (MecA1/A2) and mecA1 (mecA1-spec-F/R) (Table ).

    Article Title: HoxBlinc RNA recruits Set1/MLL complexes to activate Hox gene expression patterns and mesoderm lineage development
    Article Snippet: .. 2 µg of RNA was reverse transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs). cDNA was analyzed by real-time PCR (qRT-PCR) using the SYBR Green PCR master mix and the Bio-Rad CFX96 system (Bio-Rad). ..

    Article Title: Heterogeneity of tumor-infiltrating lymphocytes ascribed to local immune status rather than neoantigens by multi-omics analysis of glioblastoma multiforme
    Article Snippet: TCRB-specific cDNA was synthesized from 0.5 μg of total RNA using ProtoScript II Reverse Transcriptase (NEB, UK) and primer TRBCRo. .. For the first PCR step, cDNA templates were pre-amplified using a multiplex PCR 5x Master Mix (NEB, UK) with multiple Vβ region primers (TRBV1Fo-TRBV30Fo) and a Cβ region primer (TRBCRo primer).

    Article Title: Streptococcus suis contains multiple phase-variable methyltransferases that show a discrete lineage distribution
    Article Snippet: Prior to reverse transcription (RT), RNA was treated with heat-labile dsDNAse (Thermo Fisher catalogue number EN0771) according to manufacturer’s instructions, and used directly in the RT reaction. cDNA was synthesized from DNAse-treated RNA using Protoscript II Reverse Transcriptase (New England Biolabs) according to manufacturer’s instructions. .. Semi-quantitative PCR was carried out in a multiplex reaction using GoTaq DNA polymerase (Promega) in a standard PCR reaction with primers specific for 16s rRNA, and one of four Type I allele-specific primer pairs (alleles A, B, C or D; Table ).

    Isolation:

    Article Title: Overexpression of TET dioxygenases in seminomas associates with low levels of DNA methylation and hydroxymethylation
    Article Snippet: .. Alternatively, total RNA was isolated using RNA MicroPrep™ (Zymo Research) according to manufacturer’s instructions. cDNA for the quantitative RT-PCR (qRT-PCR) analysis was synthesized using Protoscript II reverse transcriptase (NEB) and random hexanucleotides (Promega). ..

    Article Title: 3′ end additions by T7 RNA polymerase are RNA self-templated, distributive and diverse in character—RNA-Seq analyses
    Article Snippet: At last, to remove ligated RNAs from the magnetic beads, we heated the reaction to 95°C for 5 min to denature the Biotin–Streptavidin interaction and isolated the supernatant from the beads. .. The reverse transcription reactions were performed in a 50 μl reaction volume containing 0.4 μM reverse transcriptase primer, 0.5 mM each deoxynucleoside triphosphate (dNTP), 10 mM dithiothreitol (DTT) and 2 μl ProtoScript® II Reverse Transcriptase (New England BioLabs) at 200 U μl−1 by incubation at 42°C for 1 h.

    Article Title: Ectopic expression of a cyanobacterial flavodoxin in creeping bentgrass impacts plant development and confers broad abiotic stress tolerance
    Article Snippet: .. RNA isolation, cDNA synthesis, qPCR, RT‐PCR and northern blot One hundred milligrams of young leaf tissues was employed for total RNA isolation using Trizol reagent (Invitrogen, Carlsbad, CA), and 2 μg of total RNAs was reverse‐transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs, GA) following manufacturer's instructions. ..

    Article Title: Loss of rps9 in Zebrafish Leads to p53-Dependent Anemia
    Article Snippet: .. Real-time quantitative PCR Total RNA was isolated using TRIzol Reagent (Life Technologies) from pool of 30 embryos according to the manufacturer’s directions. cDNA was made using ProtoScript II Reverse Transcriptase and random hexamer primers (New England Biolabs). .. Quantitative PCR was performed on Stratagene Mx3005P (Agilent Technologies) using Luna Universal qPCR Master Mix (New England Biolabs).

    Article Title: HoxBlinc RNA recruits Set1/MLL complexes to activate Hox gene expression patterns and mesoderm lineage development
    Article Snippet: Total RNA was prepared by using the RNeasy mini isolation kit according to the manufacturer’s instruction (Qiagen, MD, USA). .. 2 µg of RNA was reverse transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs). cDNA was analyzed by real-time PCR (qRT-PCR) using the SYBR Green PCR master mix and the Bio-Rad CFX96 system (Bio-Rad).

    Sequencing:

    Article Title: 3′ end additions by T7 RNA polymerase are RNA self-templated, distributive and diverse in character—RNA-Seq analyses
    Article Snippet: The reactions were incubated at 37°C for 30 min, and then stopped by heat inactivation at 65°C for 20 min. A 5′ adapter sequence, containing a barcode unique to each experiment , was then ligated to the 5′ mono-phosphorylated product by T4 RNA Ligase 1 (New England BioLabs) ( ). .. The reverse transcription reactions were performed in a 50 μl reaction volume containing 0.4 μM reverse transcriptase primer, 0.5 mM each deoxynucleoside triphosphate (dNTP), 10 mM dithiothreitol (DTT) and 2 μl ProtoScript® II Reverse Transcriptase (New England BioLabs) at 200 U μl−1 by incubation at 42°C for 1 h.

    Article Title: Global Transcriptome Analysis of Aedes aegypti Mosquitoes in Response to Zika Virus Infection
    Article Snippet: Paragraph title: Library preparations and sequencing. ... Fragmented poly(A)+ RNA samples were converted to cDNA by random primed synthesis using ProtoScript II reverse transcriptase (New England Biolabs).

    Article Title: Heterogeneity of tumor-infiltrating lymphocytes ascribed to local immune status rather than neoantigens by multi-omics analysis of glioblastoma multiforme
    Article Snippet: Paragraph title: TCR sequencing ... TCRB-specific cDNA was synthesized from 0.5 μg of total RNA using ProtoScript II Reverse Transcriptase (NEB, UK) and primer TRBCRo.

    Article Title: Identification of radiation responsive genes and transcriptome profiling via complete RNA sequencing in a stable radioresistant U87 glioblastoma model
    Article Snippet: Paragraph title: RNA library preparation and sequencing ... First strand complementary deoxyribonucleic acid (cDNA) syntheses were performed at 25° C for 10 minutes, 42° C for 15 minutes, and 70° C for 15 minutes, using random hexameres and ProtoScript II Reverse Transcriptase (New England BioLabs Inc., Ipswich, MA, USA).

    RNA Sequencing Assay:

    Article Title: 3′ end additions by T7 RNA polymerase are RNA self-templated, distributive and diverse in character—RNA-Seq analyses
    Article Snippet: Paragraph title: Library preparation for RNA-Seq ... The reverse transcription reactions were performed in a 50 μl reaction volume containing 0.4 μM reverse transcriptase primer, 0.5 mM each deoxynucleoside triphosphate (dNTP), 10 mM dithiothreitol (DTT) and 2 μl ProtoScript® II Reverse Transcriptase (New England BioLabs) at 200 U μl−1 by incubation at 42°C for 1 h.

    Article Title: Global Transcriptome Analysis of Aedes aegypti Mosquitoes in Response to Zika Virus Infection
    Article Snippet: The NEBNext Ultra II RNA library kit (New England Biolabs) was used for RNA-Seq library construction. .. Fragmented poly(A)+ RNA samples were converted to cDNA by random primed synthesis using ProtoScript II reverse transcriptase (New England Biolabs).

    Article Title: Identification of radiation responsive genes and transcriptome profiling via complete RNA sequencing in a stable radioresistant U87 glioblastoma model
    Article Snippet: RNA sequencing libraries were prepared using the TruSeq Stranded mRNA Library Prep Kit (Illumina, Inc., San Diego, CA, USA) according to the manufacturer`s protocol. .. First strand complementary deoxyribonucleic acid (cDNA) syntheses were performed at 25° C for 10 minutes, 42° C for 15 minutes, and 70° C for 15 minutes, using random hexameres and ProtoScript II Reverse Transcriptase (New England BioLabs Inc., Ipswich, MA, USA).

    Fluorescence:

    Article Title: Global Transcriptome Analysis of Aedes aegypti Mosquitoes in Response to Zika Virus Infection
    Article Snippet: All samples were quantified using a Qubit fluorescence assay (Thermo Scientific). .. Fragmented poly(A)+ RNA samples were converted to cDNA by random primed synthesis using ProtoScript II reverse transcriptase (New England Biolabs).

    Magnetic Beads:

    Article Title: 3′ end additions by T7 RNA polymerase are RNA self-templated, distributive and diverse in character—RNA-Seq analyses
    Article Snippet: At last, to remove ligated RNAs from the magnetic beads, we heated the reaction to 95°C for 5 min to denature the Biotin–Streptavidin interaction and isolated the supernatant from the beads. .. The reverse transcription reactions were performed in a 50 μl reaction volume containing 0.4 μM reverse transcriptase primer, 0.5 mM each deoxynucleoside triphosphate (dNTP), 10 mM dithiothreitol (DTT) and 2 μl ProtoScript® II Reverse Transcriptase (New England BioLabs) at 200 U μl−1 by incubation at 42°C for 1 h.

    Random Primed:

    Article Title: Global Transcriptome Analysis of Aedes aegypti Mosquitoes in Response to Zika Virus Infection
    Article Snippet: .. Fragmented poly(A)+ RNA samples were converted to cDNA by random primed synthesis using ProtoScript II reverse transcriptase (New England Biolabs). ..

    Size-exclusion Chromatography:

    Article Title: Aquaporin 5 Expression in Mouse Mammary Gland Cells Is Not Driven by Promoter Methylation
    Article Snippet: The pellet was washed two times with PBS before mRNA extraction with the Quick-RNA MiniPrep Kit (Zymo Research); cDNA synthesis was performed with the ProtoScript II Reverse Transcriptase (NEB), both according to manufacturer's instructions. .. AQP5 and GAPDH expression were quantified by qPCR with the qAQP5_human and qGAPDH_human primers, respectively, using 0.25 U Phusion HotStart II DNA Polymerase (Thermo Scientific) with an initial heating step of 98°C for 30 sec followed by 45 cycles at 99°C for 5 sec, 63°C for 15 sec, and 72°C for 5 sec (see primer sequences in Supplementary Table 1).

    Article Title: Heterogeneity of tumor-infiltrating lymphocytes ascribed to local immune status rather than neoantigens by multi-omics analysis of glioblastoma multiforme
    Article Snippet: TCRB-specific cDNA was synthesized from 0.5 μg of total RNA using ProtoScript II Reverse Transcriptase (NEB, UK) and primer TRBCRo. .. The PCR protocol was 95 °C for 3 min, 10 cycles of 95 °C for 30 sec, 60 °C for 2 min, and 68 °C for 1 min; and then 68 °C for 5 min. Two microliters of the PCR product, the TRBV1Fi-TRBV30Fi, TRBCRi primer and H2O were added to the NEB multiplex PCR master mix for the second round of PCR without changing the cycling conditions.

    Mouse Assay:

    Article Title: Aquaporin 5 Expression in Mouse Mammary Gland Cells Is Not Driven by Promoter Methylation
    Article Snippet: The pellet was washed two times with PBS before mRNA extraction with the Quick-RNA MiniPrep Kit (Zymo Research); cDNA synthesis was performed with the ProtoScript II Reverse Transcriptase (NEB), both according to manufacturer's instructions. .. Primers were designed to anneal in conserved regions between humans and mice; therefore, the same plasmids could be used as standards (107 , 106 , 105 , 104 , and 103 molecules/reaction) for measuring murine or human absolute AQP5 mRNA levels.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Ectopic expression of a cyanobacterial flavodoxin in creeping bentgrass impacts plant development and confers broad abiotic stress tolerance
    Article Snippet: .. RNA isolation, cDNA synthesis, qPCR, RT‐PCR and northern blot One hundred milligrams of young leaf tissues was employed for total RNA isolation using Trizol reagent (Invitrogen, Carlsbad, CA), and 2 μg of total RNAs was reverse‐transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs, GA) following manufacturer's instructions. ..

    Article Title: A novel hybrid SCCmec-mecC region in Staphylococcus sciuri
    Article Snippet: Paragraph title: Transcriptional analysis of mec gene expression by RT–PCR ... After an additional DNAse step using RQ1 RNase-Free DNase (Promega, UK), cDNA was synthesized using ProtoScript® II Reverse Transcriptase (NEB, UK) and a Random Hexamer primer (Fisher Scientific, UK) following the manufacturer's standard protocol.

    Article Title: Streptococcus suis contains multiple phase-variable methyltransferases that show a discrete lineage distribution
    Article Snippet: Paragraph title: RNA preparation and semi-quantitative RT-PCR ... Prior to reverse transcription (RT), RNA was treated with heat-labile dsDNAse (Thermo Fisher catalogue number EN0771) according to manufacturer’s instructions, and used directly in the RT reaction. cDNA was synthesized from DNAse-treated RNA using Protoscript II Reverse Transcriptase (New England Biolabs) according to manufacturer’s instructions.

    Nested PCR:

    Article Title: Heterogeneity of tumor-infiltrating lymphocytes ascribed to local immune status rather than neoantigens by multi-omics analysis of glioblastoma multiforme
    Article Snippet: TCR sequencing The TCR sequences were obtained via a series of three nested PCR reactions using the primers as described in ref. (Table ) with a slightly modified protocol of the ARM-PCR procedure , . .. TCRB-specific cDNA was synthesized from 0.5 μg of total RNA using ProtoScript II Reverse Transcriptase (NEB, UK) and primer TRBCRo.

    Purification:

    Article Title: Identification of radiation responsive genes and transcriptome profiling via complete RNA sequencing in a stable radioresistant U87 glioblastoma model
    Article Snippet: Briefly, polyA mRNA from an input of 500 ng high-quality total RNA (RINe > 8) was purified and fragmented. .. First strand complementary deoxyribonucleic acid (cDNA) syntheses were performed at 25° C for 10 minutes, 42° C for 15 minutes, and 70° C for 15 minutes, using random hexameres and ProtoScript II Reverse Transcriptase (New England BioLabs Inc., Ipswich, MA, USA).

    Article Title: Design, synthesis, and biological evaluation of inhibitors of the NADPH oxidase, Nox4
    Article Snippet: Total RNA samples (2 μg), purified from cultured MASMs, using the RNeasy Plus kit (Qiagen, Chatsworth, CA), were used to prepare cDNA. .. Reactions were carried out at 42 °C, using random 15-mer primers and ProtoScript II reverse transcriptase (New England Biolabs, Beverly, MA).

    Chromatin Immunoprecipitation:

    Article Title: Global Transcriptome Analysis of Aedes aegypti Mosquitoes in Response to Zika Virus Infection
    Article Snippet: Total RNA quality was assessed using an RNA 6000 chip on an Agilent 2100 Bioanalyzer (Agilent Technologies). .. Fragmented poly(A)+ RNA samples were converted to cDNA by random primed synthesis using ProtoScript II reverse transcriptase (New England Biolabs).

    Plasmid Preparation:

    Article Title: In Vitro Study on Anti-Hepatitis C Virus Activity of Spatholobus suberectus Dunn
    Article Snippet: One microgram of total RNA was reverse transcribed into cDNA with ProtoScript® II Reverse Transcriptase (New England Biolabs, Ipswich, MA, USA). qPCR was performed in triplicates for all samples using the Mx 3005P qPCR system (Agilent, Santa Clara, CA, USA) with SYBR Master Mix (Bio-Rad, Hercules, CA, USA). .. Absolute copy number of HCV RNA was determined with the standard curve generated by using the linearized pFK-I389/NS3-3′ plasmid bearing the HCV genome and 18S rRNA was included as a control.

    Real-time Polymerase Chain Reaction:

    Article Title: In Vitro Study on Anti-Hepatitis C Virus Activity of Spatholobus suberectus Dunn
    Article Snippet: .. One microgram of total RNA was reverse transcribed into cDNA with ProtoScript® II Reverse Transcriptase (New England Biolabs, Ipswich, MA, USA). qPCR was performed in triplicates for all samples using the Mx 3005P qPCR system (Agilent, Santa Clara, CA, USA) with SYBR Master Mix (Bio-Rad, Hercules, CA, USA). .. Absolute copy number of HCV RNA was determined with the standard curve generated by using the linearized pFK-I389/NS3-3′ plasmid bearing the HCV genome and 18S rRNA was included as a control.

    Article Title: Aquaporin 5 Expression in Mouse Mammary Gland Cells Is Not Driven by Promoter Methylation
    Article Snippet: The pellet was washed two times with PBS before mRNA extraction with the Quick-RNA MiniPrep Kit (Zymo Research); cDNA synthesis was performed with the ProtoScript II Reverse Transcriptase (NEB), both according to manufacturer's instructions. .. AQP5 and GAPDH expression were quantified by qPCR with the qAQP5_human and qGAPDH_human primers, respectively, using 0.25 U Phusion HotStart II DNA Polymerase (Thermo Scientific) with an initial heating step of 98°C for 30 sec followed by 45 cycles at 99°C for 5 sec, 63°C for 15 sec, and 72°C for 5 sec (see primer sequences in Supplementary Table 1).

    Article Title: Ectopic expression of a cyanobacterial flavodoxin in creeping bentgrass impacts plant development and confers broad abiotic stress tolerance
    Article Snippet: .. RNA isolation, cDNA synthesis, qPCR, RT‐PCR and northern blot One hundred milligrams of young leaf tissues was employed for total RNA isolation using Trizol reagent (Invitrogen, Carlsbad, CA), and 2 μg of total RNAs was reverse‐transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs, GA) following manufacturer's instructions. ..

    Article Title: Loss of rps9 in Zebrafish Leads to p53-Dependent Anemia
    Article Snippet: .. Real-time quantitative PCR Total RNA was isolated using TRIzol Reagent (Life Technologies) from pool of 30 embryos according to the manufacturer’s directions. cDNA was made using ProtoScript II Reverse Transcriptase and random hexamer primers (New England Biolabs). .. Quantitative PCR was performed on Stratagene Mx3005P (Agilent Technologies) using Luna Universal qPCR Master Mix (New England Biolabs).

    Article Title: HoxBlinc RNA recruits Set1/MLL complexes to activate Hox gene expression patterns and mesoderm lineage development
    Article Snippet: .. 2 µg of RNA was reverse transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs). cDNA was analyzed by real-time PCR (qRT-PCR) using the SYBR Green PCR master mix and the Bio-Rad CFX96 system (Bio-Rad). ..

    Article Title: Design, synthesis, and biological evaluation of inhibitors of the NADPH oxidase, Nox4
    Article Snippet: Paragraph title: 4.3.5 Reverse transcription and quantitative PCR ... Reactions were carried out at 42 °C, using random 15-mer primers and ProtoScript II reverse transcriptase (New England Biolabs, Beverly, MA).

    Multiplex Assay:

    Article Title: Heterogeneity of tumor-infiltrating lymphocytes ascribed to local immune status rather than neoantigens by multi-omics analysis of glioblastoma multiforme
    Article Snippet: TCRB-specific cDNA was synthesized from 0.5 μg of total RNA using ProtoScript II Reverse Transcriptase (NEB, UK) and primer TRBCRo. .. For the first PCR step, cDNA templates were pre-amplified using a multiplex PCR 5x Master Mix (NEB, UK) with multiple Vβ region primers (TRBV1Fo-TRBV30Fo) and a Cβ region primer (TRBCRo primer).

    Article Title: Streptococcus suis contains multiple phase-variable methyltransferases that show a discrete lineage distribution
    Article Snippet: Prior to reverse transcription (RT), RNA was treated with heat-labile dsDNAse (Thermo Fisher catalogue number EN0771) according to manufacturer’s instructions, and used directly in the RT reaction. cDNA was synthesized from DNAse-treated RNA using Protoscript II Reverse Transcriptase (New England Biolabs) according to manufacturer’s instructions. .. Semi-quantitative PCR was carried out in a multiplex reaction using GoTaq DNA polymerase (Promega) in a standard PCR reaction with primers specific for 16s rRNA, and one of four Type I allele-specific primer pairs (alleles A, B, C or D; Table ).

    Agarose Gel Electrophoresis:

    Article Title: Ectopic expression of a cyanobacterial flavodoxin in creeping bentgrass impacts plant development and confers broad abiotic stress tolerance
    Article Snippet: RNA isolation, cDNA synthesis, qPCR, RT‐PCR and northern blot One hundred milligrams of young leaf tissues was employed for total RNA isolation using Trizol reagent (Invitrogen, Carlsbad, CA), and 2 μg of total RNAs was reverse‐transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs, GA) following manufacturer's instructions. .. PCR was conducted with the following program: an initial DNA polymerase activation at 95 °C for 180 s followed by 40 cycles of 95 °C for 30 s, 60 °C for 20 s and 72 °C for 20 s. Finally, a melting curve was performed, and the PCR products were checked with 2% agarose gel in 0.5× TBE buffer with ethidium bromide.

    Random Hexamer Labeling:

    Article Title: A novel hybrid SCCmec-mecC region in Staphylococcus sciuri
    Article Snippet: .. After an additional DNAse step using RQ1 RNase-Free DNase (Promega, UK), cDNA was synthesized using ProtoScript® II Reverse Transcriptase (NEB, UK) and a Random Hexamer primer (Fisher Scientific, UK) following the manufacturer's standard protocol. .. Controls without reverse transcriptase were generated for all samples and showed no amplification in the subsequent PCRs. cDNA was used undiluted in a standard PCR for the detection of mecC (mecC-Uni-F/R), mecA (MecA1/A2) and mecA1 (mecA1-spec-F/R) (Table ).

    Article Title: Loss of rps9 in Zebrafish Leads to p53-Dependent Anemia
    Article Snippet: .. Real-time quantitative PCR Total RNA was isolated using TRIzol Reagent (Life Technologies) from pool of 30 embryos according to the manufacturer’s directions. cDNA was made using ProtoScript II Reverse Transcriptase and random hexamer primers (New England Biolabs). .. Quantitative PCR was performed on Stratagene Mx3005P (Agilent Technologies) using Luna Universal qPCR Master Mix (New England Biolabs).

    Spectrophotometry:

    Article Title: Identification of radiation responsive genes and transcriptome profiling via complete RNA sequencing in a stable radioresistant U87 glioblastoma model
    Article Snippet: The RNA concentration was measured with a NanoDrop 2000c spectrophotometer (Thermo Scientific Inc., Waltham, MA, USA). .. First strand complementary deoxyribonucleic acid (cDNA) syntheses were performed at 25° C for 10 minutes, 42° C for 15 minutes, and 70° C for 15 minutes, using random hexameres and ProtoScript II Reverse Transcriptase (New England BioLabs Inc., Ipswich, MA, USA).

    Activation Assay:

    Article Title: Ectopic expression of a cyanobacterial flavodoxin in creeping bentgrass impacts plant development and confers broad abiotic stress tolerance
    Article Snippet: RNA isolation, cDNA synthesis, qPCR, RT‐PCR and northern blot One hundred milligrams of young leaf tissues was employed for total RNA isolation using Trizol reagent (Invitrogen, Carlsbad, CA), and 2 μg of total RNAs was reverse‐transcribed using ProtoScript® II Reverse Transcriptase (New England Biolabs, GA) following manufacturer's instructions. .. PCR was conducted with the following program: an initial DNA polymerase activation at 95 °C for 180 s followed by 40 cycles of 95 °C for 30 s, 60 °C for 20 s and 72 °C for 20 s. Finally, a melting curve was performed, and the PCR products were checked with 2% agarose gel in 0.5× TBE buffer with ethidium bromide.

    Concentration Assay:

    Article Title: Superinfection with Woodchuck Hepatitis Virus Strain WHVNY of Livers Chronically Infected with Strain WHV7
    Article Snippet: .. The cDNA synthesis reaction was started by adding 200 U of ProtoScript II reverse transcriptase (RT; New England BioLabs [NEB]), 2 μl of 10× ProtoScript II buffer, and dithiothreitol (DTT) (to a final concentration of 10 mM). .. After incubation for 1 h at 42°C, an additional 200 U of ProtoScript II reverse transcriptase was added to the reaction mixture.

    Article Title: Identification of radiation responsive genes and transcriptome profiling via complete RNA sequencing in a stable radioresistant U87 glioblastoma model
    Article Snippet: The RNA concentration was measured with a NanoDrop 2000c spectrophotometer (Thermo Scientific Inc., Waltham, MA, USA). .. First strand complementary deoxyribonucleic acid (cDNA) syntheses were performed at 25° C for 10 minutes, 42° C for 15 minutes, and 70° C for 15 minutes, using random hexameres and ProtoScript II Reverse Transcriptase (New England BioLabs Inc., Ipswich, MA, USA).

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    New England Biolabs protoscript ii reverse transcriptase
    Protoscript Ii Reverse Transcriptase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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