Total RNA was purified from zebrafish embryos (40 embryos/condition), human CD34+ cells (500,000 cells/condition), or mononuclear UCB (one million cells/condition) using the RNAqueous Micro kit (Life Technologies) followed by Turbo DNase-I treatment (Life Technologies). For sorted populations, cDNA amplification was performed using the NuGEN Ovation Pico WTA2 system. For embryos, 1 µg of total RNA was used to generate cDNA using Superscript III First Strand Synthesis Supermix (Life Technologies). Superscript VILO mastermix was used for human RNA samples (<1 µg of RNA). Quantitative real-time PCR was performed using SYBR Green PCR Master Mix (Life Technologies) on a Bio-Rad CFX384 Touch. Samples were run in triplicate with more than three biological replicates using published primers (Supplemental Experimental Procedures). Data analysis was performed using Real PCR Miner (http://ewindup.info/miner/ ).
Turbo dnase 1 treatment
Manufactured by Thermo Fisher Scientific
Turbo DNase-I treatment is a laboratory product designed to remove DNA from RNA samples. It functions by utilizing the DNase-I enzyme to degrade DNA, allowing for the isolation and purification of RNA.
Automatically generated - may contain errors
Lab products found in correlation
Rnaqueous micro kit, by Thermo Fisher Scientific (2 mentions)
Superscript 3 first strand synthesis supermix, by Thermo Fisher Scientific (2 mentions)
Cfx384 touch, by Bio-Rad (2 mentions)
Sybr green pcr master mix, by Thermo Fisher Scientific (2 mentions)
Nugen ovation pico wta2 system, by Tecan (2 mentions)
2 protocols using turbo dnase 1 treatment
1
Quantitative Real-Time PCR Across Cell Types
2
Quantitative Real-Time PCR Across Cell Types
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Total RNA was purified from zebrafish embryos (40 embryos/condition), human CD34+ cells (500,000 cells/condition), or mononuclear UCB (one million cells/condition) using the RNAqueous Micro kit (Life Technologies) followed by Turbo DNase-I treatment (Life Technologies). For sorted populations, cDNA amplification was performed using the NuGEN Ovation Pico WTA2 system. For embryos, 1 µg of total RNA was used to generate cDNA using Superscript III First Strand Synthesis Supermix (Life Technologies). Superscript VILO mastermix was used for human RNA samples (<1 µg of RNA). Quantitative real-time PCR was performed using SYBR Green PCR Master Mix (Life Technologies) on a Bio-Rad CFX384 Touch. Samples were run in triplicate with more than three biological replicates using published primers (Supplemental Experimental Procedures). Data analysis was performed using Real PCR Miner (http://ewindup.info/miner/ ).
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