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iscript reverse transcriptase kit  (Bio-Rad)


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    Structured Review

    Bio-Rad iscript reverse transcriptase kit
    (A–C) Dose–response curves using double-bubble primer mix #8 and VIC-TqM probe #12 and decimal dilutions of SARS-CoV-2 synthetic RNA N. (A) Real-time amplification plot. C t values: 8 × 10 5 copies, 22.82; 8 × 10 4 copies, 26.27; 8 × 10 3 copies, 30.55; 8 × 10 2 copies, 32.51. (B) 5% agarose gel of the PCR amplification products. Lane 1: ultra-low-range ladder; lanes 2–5: 139-bp amplicons of 8 × 10 5 to 8 × 10 2 RNA copies per tube. Also included in the PCR reaction mix were 2× TqM buffer without UDP and <t>iScript</t> reverse-transcriptase. (C) Efficiency plot of the PCR reaction depicted in panel A. Slope = -3.3359 represents PCR reaction efficiency of 99.42%; R 2 = 0.9816. (D) One-tube RT-qPCR using double-bubble primer mix #10 from region 2 using SARS-CoV-2 synthetic RNA (2 × 10 4 copies per tube) and FAM-TqM probe #13. RT-qPCR was performed with standard conditions.
    Iscript Reverse Transcriptase Kit, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 48597 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/goscript+reverse+transcriptase+rt-pcr+kit/iScript+cDNA+Synthesis+Kit/pmc08544493-38-3-19
    Average 99 stars, based on 48597 article reviews
    iscript reverse transcriptase kit - by Bioz Stars, 2026-11
    99/100 stars

    Images

    1) Product Images from "Improved SARS-CoV-2 PCR detection and genotyping with double-bubble primers"

    Article Title: Improved SARS-CoV-2 PCR detection and genotyping with double-bubble primers

    Journal: Biotechniques

    doi: 10.2144/btn-2021-0063

    (A–C) Dose–response curves using double-bubble primer mix #8 and VIC-TqM probe #12 and decimal dilutions of SARS-CoV-2 synthetic RNA N. (A) Real-time amplification plot. C t values: 8 × 10 5 copies, 22.82; 8 × 10 4 copies, 26.27; 8 × 10 3 copies, 30.55; 8 × 10 2 copies, 32.51. (B) 5% agarose gel of the PCR amplification products. Lane 1: ultra-low-range ladder; lanes 2–5: 139-bp amplicons of 8 × 10 5 to 8 × 10 2 RNA copies per tube. Also included in the PCR reaction mix were 2× TqM buffer without UDP and iScript reverse-transcriptase. (C) Efficiency plot of the PCR reaction depicted in panel A. Slope = -3.3359 represents PCR reaction efficiency of 99.42%; R 2 = 0.9816. (D) One-tube RT-qPCR using double-bubble primer mix #10 from region 2 using SARS-CoV-2 synthetic RNA (2 × 10 4 copies per tube) and FAM-TqM probe #13. RT-qPCR was performed with standard conditions.
    Figure Legend Snippet: (A–C) Dose–response curves using double-bubble primer mix #8 and VIC-TqM probe #12 and decimal dilutions of SARS-CoV-2 synthetic RNA N. (A) Real-time amplification plot. C t values: 8 × 10 5 copies, 22.82; 8 × 10 4 copies, 26.27; 8 × 10 3 copies, 30.55; 8 × 10 2 copies, 32.51. (B) 5% agarose gel of the PCR amplification products. Lane 1: ultra-low-range ladder; lanes 2–5: 139-bp amplicons of 8 × 10 5 to 8 × 10 2 RNA copies per tube. Also included in the PCR reaction mix were 2× TqM buffer without UDP and iScript reverse-transcriptase. (C) Efficiency plot of the PCR reaction depicted in panel A. Slope = -3.3359 represents PCR reaction efficiency of 99.42%; R 2 = 0.9816. (D) One-tube RT-qPCR using double-bubble primer mix #10 from region 2 using SARS-CoV-2 synthetic RNA (2 × 10 4 copies per tube) and FAM-TqM probe #13. RT-qPCR was performed with standard conditions.

    Techniques Used: Amplification, Agarose Gel Electrophoresis, Reverse Transcription, Quantitative RT-PCR

    (A) qPCR using reverse-transcribed cDNA template and double-bubble (D-B) primer mix #10, FAM-TqM probe #13, TqM fast kit+ uracil-DNA glycosylase (UDG) assayed in duplicates. Insert depicts 5% agarose gel; lane 1: ultra-low-range (ULR) ladder; lanes 2 & 3: template cDNA; lanes 4 & 5: non-template control (NTC). C t average values: cDNA, 28.60; NTC, undetermined. (B) One-tube RT-qPCR using SARS-CoV-2 synthetic RNA N template and D-B primer mix #8, VIC-TqM probe #12, TqM fast kit without UDG iScript reverse transcriptase, assayed in duplicate. Insert depicts 5% agarose gel; lane 1: ULR ladder; lanes 2 and 3: template RNA; lanes 4 and 5: NTC. C t average values: RNA, 24.63, NTC, undetermined. (C) Duplex qPCR using SARS-CoV-2 synthetic RNA N template and TqM fast kit with UDG. C t values: primer 8, 26.13; primer 10, 32.19; duplex primers 8 + 10, 26.36 and 30.27, respectively; NTC, undetermined. (D) 5% agarose gel of samples shown in panel C. Lane 1: ULR ladder; lane 2: primer D-B mix #8 (amplicon 139 bp); lane 3: D-B primer mix #10 (amplicon 158 bp); lane 4: duplex of both primers #8 and #10 (amplicons 139 and 158 bp). Arrows (C & D) show duplex amplifications in the same tube with D-B primers #8 and #10. PCR was performed using fast conditions with 40 (A, C & D) and 30 (B) cycles.
    Figure Legend Snippet: (A) qPCR using reverse-transcribed cDNA template and double-bubble (D-B) primer mix #10, FAM-TqM probe #13, TqM fast kit+ uracil-DNA glycosylase (UDG) assayed in duplicates. Insert depicts 5% agarose gel; lane 1: ultra-low-range (ULR) ladder; lanes 2 & 3: template cDNA; lanes 4 & 5: non-template control (NTC). C t average values: cDNA, 28.60; NTC, undetermined. (B) One-tube RT-qPCR using SARS-CoV-2 synthetic RNA N template and D-B primer mix #8, VIC-TqM probe #12, TqM fast kit without UDG iScript reverse transcriptase, assayed in duplicate. Insert depicts 5% agarose gel; lane 1: ULR ladder; lanes 2 and 3: template RNA; lanes 4 and 5: NTC. C t average values: RNA, 24.63, NTC, undetermined. (C) Duplex qPCR using SARS-CoV-2 synthetic RNA N template and TqM fast kit with UDG. C t values: primer 8, 26.13; primer 10, 32.19; duplex primers 8 + 10, 26.36 and 30.27, respectively; NTC, undetermined. (D) 5% agarose gel of samples shown in panel C. Lane 1: ULR ladder; lane 2: primer D-B mix #8 (amplicon 139 bp); lane 3: D-B primer mix #10 (amplicon 158 bp); lane 4: duplex of both primers #8 and #10 (amplicons 139 and 158 bp). Arrows (C & D) show duplex amplifications in the same tube with D-B primers #8 and #10. PCR was performed using fast conditions with 40 (A, C & D) and 30 (B) cycles.

    Techniques Used: Reverse Transcription, Agarose Gel Electrophoresis, Control, Quantitative RT-PCR, Amplification

    The reaction mixture contained hot-start double-bubble (D-B) primer mix #4, VIC-TqM probe #12, cost-effective non-hot-start Taq polymerase (FroggaBio), reverse transcriptase (iScript), synthetic SARS-CoV-2 RNA gene N as template, added dNTPs and ROX dye for internal calibration. The reaction was assembled at room temperature and subjected to real time RT-qPCR using the following fast conditions: 42°C, 15 min; 95°C,1 min; 30 cycles of 95°C, 1 s → 70°C, 20 s. The amplification plot and (insert) 5% agarose gel depict the amplification of SARS-CoV-2 RNA extracted from the nasopharyngeal swabs of patient S1 (lane 2) and negative patient N1 (lane 3), no-template controls (lanes 4 and 5) and SARS-CoV-2 gene N synthetic RNA as positive control (lane 6). Note the amplification in the real-time plot and the 115-bp band in the 5% agarose gel with SARS-CoV-2 virus or synthetic RNA (lanes 2 and 6), with no amplifications in the negative control N1 (lane 3) or NTC (lanes 4 and 5). Lane 1: ultra-low-range ladder. C t values: positive patient S1: 23.95; synthetic RNA: 23.84; negative patient N1 and NTC: undetermined.
    Figure Legend Snippet: The reaction mixture contained hot-start double-bubble (D-B) primer mix #4, VIC-TqM probe #12, cost-effective non-hot-start Taq polymerase (FroggaBio), reverse transcriptase (iScript), synthetic SARS-CoV-2 RNA gene N as template, added dNTPs and ROX dye for internal calibration. The reaction was assembled at room temperature and subjected to real time RT-qPCR using the following fast conditions: 42°C, 15 min; 95°C,1 min; 30 cycles of 95°C, 1 s → 70°C, 20 s. The amplification plot and (insert) 5% agarose gel depict the amplification of SARS-CoV-2 RNA extracted from the nasopharyngeal swabs of patient S1 (lane 2) and negative patient N1 (lane 3), no-template controls (lanes 4 and 5) and SARS-CoV-2 gene N synthetic RNA as positive control (lane 6). Note the amplification in the real-time plot and the 115-bp band in the 5% agarose gel with SARS-CoV-2 virus or synthetic RNA (lanes 2 and 6), with no amplifications in the negative control N1 (lane 3) or NTC (lanes 4 and 5). Lane 1: ultra-low-range ladder. C t values: positive patient S1: 23.95; synthetic RNA: 23.84; negative patient N1 and NTC: undetermined.

    Techniques Used: Reverse Transcription, Quantitative RT-PCR, Amplification, Agarose Gel Electrophoresis, Positive Control, Virus, Negative Control

    Related Articles

    cDNA Synthesis:

    Article Title: Pharmacological melatonin induces mitochondrial dysfunction and alters extracellular nucleotide metabolism and CD39/CD73 expression in human melanoma cells
    Article Snippet: Total RNA was extracted using TRIzol reagent (Invitrogen), and concentration and purity were determined by spectrophotometry (Varioskan LUX, Thermo Fisher Scientific) at 260/280 nm. .. RNA samples were treated with DNase, and cDNA synthesis was performed using the MultiScript cDNA Synthesis Kit (Bio-Rad), according to the manufacturer's instructions. .. qPCR reactions were carried out using 3 μL of cDNA and PowerUp SYBR Green PCR Master Mix (Thermo Scientific) in a final volume of 10 μL, under standard thermal cycling conditions.

    Article Title: Urinary KIM-1 Transcript Detectability in Workers Chronically Exposed to Arsenic and Antimony in an Industrial Area of Central Chile: A Cross-Sectional Pilot Study.
    Article Snippet: Background: Chronic arsenic (As) and antimony (Sb) exposure causes renal toxicity in experimental models, but early molecular indicators of tubular stress in humans remain uncharacterised.. Jo ur na l P re -p ro of Methods: We assessed 12 male workers from Puchuncaví, Chile.. Urinary and blood As and Sb were quantified by hydride-generation atomic fluorescence spectrometry (HG-AFS).

    Article Title: Hesperidin Attenuates Vincristine‐Induced Nephrotoxicity via Nrf2/HO‐1 and PI3K/Akt/mTOR Signaling
    Article Snippet: The RNA concentrations were standardized across samples, and the RNA purity and quantity were assessed using a NanoDrop spectrophotometer (BioTek Instruments, EPOCH, USA). .. In accordance with the manufacturer's instructions, the iScript cDNA Synthesis Kit (Bio‐Rad, USA) was employed to generate complementary DNA (cDNA). .. The qRT‐PCR was conducted using a Rotor‐Gene Q system (Qiagen, Germany) and iTaq Universal SYBR Green Supermix (Bio‐Rad, USA).

    Article Title: Diarylheptanoids oxyphyllacinol, yakuchinone A, and yakuchinone B attenuate 12-O-tetradecanoylphorbol-13-acetate-induced acute skin inflammation in mice
    Article Snippet: Diarylheptanoids from Alpinia oxyphylla are potential anti-inflammatory compounds, but existing evidence is limited and has largely focused on YAK-A and YAK-B, with little direct comparative evaluation of structurally related diarylheptanoids in inflammatory models.. Here, we compared the protective effects of oxyphyllacinol (OXY), yakuchinone A (YAK-A), and yakuchinone B (YAK-B) in a 12-O-tetradecanoylphorbol-13-acetate (TPA)induced mouse ear inflammation model. Topical pretreatment with OXY, YAK-A, or YAK-B significantly attenuated TPA-induced ear edema and tissue thickening, with YAK-B showing the greatest reduction in edema under the experimental conditions examined.. These compounds differentially regulated inflammatory cytokine expression, with OXY showing more consistent inhibition of IL-1β and IL-6 at the protein level.

    Article Title: Foundational characterization of tomato fruit RALF peptides reveals structural and functional specialization within the SlRALF family.
    Article Snippet: RNA integrity was verified by agarose gel electrophoresis, and RNA purity was assessed using a NanoDrop spectrophotometer (Thermo Fisher Scientific, USA). .. For cDNA synthesis, 1 μg of total RNA was used for cDNA synthesis using oligo(dT) primers and the iScript cDNA Synthesis Kit (Bio-Rad) following the manufactureŕs instructions. .. RT-qPCR was performed using SsoFast EvaGreen Supermix (Bio-Rad) on a CFX384 Real-Time PCR Detection System (Bio-Rad).

    Article Title: Furan fatty acids reprogram hepatic lipid metabolism via acyl-CoA conjugates in obesity.
    Article Snippet: .. One microgram of RNA was reverse transcribed in a total volume of 20 μL using the iScriptTM cDNA Synthesis Kit (Bio- Rad). .. Quantitative PCR was performed using TaqManTM Fast Advanced Master Mix (Applied Biosystems) and TaqManTM Gene Expression Assays (Applied Biosystems).

    Article Title: Modelling herpes simplex virus type 2 and Zika virus replication in mouse vaginal organoids and human vaginal spheroids.
    Article Snippet: Organoid pellets were resuspended in 300 μL of RNA lysis buffer (RNeasy Mini Kit, Qiagen Cat. No. 74104), and RNA was extracted according to the manufacturer’s instructions. .. 1 μg of RNA was reverse transcribed to cDNA using the iScript cDNA Synthesis Kit (BioRad Cat No. 1708891). .. RT-qPCR was performed using QuantStudio 3 (Thermo Fisher) and PowerTrack SYBR Green Master Mix (Thermo Fisher Cat No. A46110).

    Article Title: Arginine Supplementation for Juvenile Florida Pompano ( Trachinotus carolinus ) in Practical Plant‐Based Diets
    Article Snippet: .. Complementary DNA was synthesized using the iScript cDNA Synthesis Kit (Bio‐Rad Laboratories, Hercules, California, USA). ..

    Synthesized:

    Article Title: Urinary KIM-1 Transcript Detectability in Workers Chronically Exposed to Arsenic and Antimony in an Industrial Area of Central Chile: A Cross-Sectional Pilot Study.
    Article Snippet: Background: Chronic arsenic (As) and antimony (Sb) exposure causes renal toxicity in experimental models, but early molecular indicators of tubular stress in humans remain uncharacterised.. Jo ur na l P re -p ro of Methods: We assessed 12 male workers from Puchuncaví, Chile.. Urinary and blood As and Sb were quantified by hydride-generation atomic fluorescence spectrometry (HG-AFS).

    Article Title: Diarylheptanoids oxyphyllacinol, yakuchinone A, and yakuchinone B attenuate 12-O-tetradecanoylphorbol-13-acetate-induced acute skin inflammation in mice
    Article Snippet: Diarylheptanoids from Alpinia oxyphylla are potential anti-inflammatory compounds, but existing evidence is limited and has largely focused on YAK-A and YAK-B, with little direct comparative evaluation of structurally related diarylheptanoids in inflammatory models.. Here, we compared the protective effects of oxyphyllacinol (OXY), yakuchinone A (YAK-A), and yakuchinone B (YAK-B) in a 12-O-tetradecanoylphorbol-13-acetate (TPA)induced mouse ear inflammation model. Topical pretreatment with OXY, YAK-A, or YAK-B significantly attenuated TPA-induced ear edema and tissue thickening, with YAK-B showing the greatest reduction in edema under the experimental conditions examined.. These compounds differentially regulated inflammatory cytokine expression, with OXY showing more consistent inhibition of IL-1β and IL-6 at the protein level.

    Article Title: Arginine Supplementation for Juvenile Florida Pompano ( Trachinotus carolinus ) in Practical Plant‐Based Diets
    Article Snippet: .. Complementary DNA was synthesized using the iScript cDNA Synthesis Kit (Bio‐Rad Laboratories, Hercules, California, USA). ..

    Reverse Transcription:

    Article Title: Furan fatty acids reprogram hepatic lipid metabolism via acyl-CoA conjugates in obesity.
    Article Snippet: .. One microgram of RNA was reverse transcribed in a total volume of 20 μL using the iScriptTM cDNA Synthesis Kit (Bio- Rad). .. Quantitative PCR was performed using TaqManTM Fast Advanced Master Mix (Applied Biosystems) and TaqManTM Gene Expression Assays (Applied Biosystems).

    Article Title: Modelling herpes simplex virus type 2 and Zika virus replication in mouse vaginal organoids and human vaginal spheroids.
    Article Snippet: Organoid pellets were resuspended in 300 μL of RNA lysis buffer (RNeasy Mini Kit, Qiagen Cat. No. 74104), and RNA was extracted according to the manufacturer’s instructions. .. 1 μg of RNA was reverse transcribed to cDNA using the iScript cDNA Synthesis Kit (BioRad Cat No. 1708891). .. RT-qPCR was performed using QuantStudio 3 (Thermo Fisher) and PowerTrack SYBR Green Master Mix (Thermo Fisher Cat No. A46110).



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