Total RNA (250ng) samples from individual mice (n = 5 per experimental and control group) and from Universal Mouse Reference total RNA (UMRR; Agilent Technologies, Inc., Mississauga, ON, Canada) were used to synthesise double-stranded complementary DNA (cDNA), which was then used to synthesise Cyanine-labelled cRNAs using Quick Amp Labeling Kit (Agilent Technologies Inc., Mississauga, ON, Canada) according to the manufacturer’s instructions. cRNAs from experimental (control and TiO2NPs-exposed) groups were labelled with cyanine 5-CTP and reference cRNAs were labelled with cyanine 3-CTP using a T7 RNA polymerase in vitro transcription kit (Agilent Technologies Inc., Mississauga, ON, Canada) and purified using RNeasy Mini kits (Qiagen, Mississauga, ON, Canada). An equimolar amount of reference cRNA was mixed with each experimental cRNA sample and was hybridised to Agilent mouse 4×44k oligonucleotide microarrays (Agilent Technologies Inc., Mississauga, ON, Canada) for 17h in a hybridisation chamber at 65°C with a rotation speed of 10rpm. At the end of hybridization, arrays were scanned on an Agilent G2505B scanner according to manufacturer’s protocols (Agilent Technologies Inc., Mississauga, ON, Canada). Gene expression data from the scanned images were extracted using Agilent Feature Extraction software version 9.5.3.1.
T7 rna polymerase in vitro transcription kit
Manufactured by Agilent Technologies
Sourced in Canada
The T7 RNA polymerase in vitro transcription kit is a laboratory tool used for the synthesis of RNA molecules. The kit contains the T7 RNA polymerase enzyme, which is responsible for the transcription of DNA templates into RNA, as well as the necessary buffers and reagents to facilitate the in vitro transcription process.
Automatically generated - may contain errors
Lab products found in correlation
Rneasy mini kit, by Qiagen (3 mentions)
Quick amp labeling kit, by Agilent Technologies (2 mentions)
Universal mouse reference total rna umrr, by Agilent Technologies (2 mentions)
Feature extraction software version 9, by Agilent Technologies (2 mentions)
G2505b scanner, by Agilent Technologies (2 mentions)
Quick amp labelling kit, by Agilent Technologies (1 mentions)
3 protocols using t7 rna polymerase in vitro transcription kit
1
Mouse total RNA microarray analysis
2
Microarray-based Gene Expression Analysis
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Double-stranded cDNA was synthesized from 250 ng total RNA of each mouse and from Universal Mouse Reference total RNA (UMRR) (Agilent Technologies, Mississauga, Canada). Cyanine-labeled cRNAs were made from the cDNA with the Quick Amp Labeling Kit (Agilent Technologies, Mississauga, Canada), and then cRNAs from each treatment group were labeled with Cyanine 5-CTP. Reference cRNAs were labeled with Cyanine 3-CTP in a T7 RNA polymerase in vitro transcription kit (Agilent Technologies, Mississauga, Canada). Purification was done with an RNeasy Mini kits (Qiagen, Mississauga, Canada). Each experimental cRNA sample was mixed with an equimolar amount of reference cRNA, and then hybridized to Agilent mouse 8 × 60 k oligonucleotide microarrays (Agilent Technologies Inc., Mississauga, Canada) for 17 h at 65 °C in a hybridization chamber rotating at 10 rpm. Next, the arrays were read in an Agilent G2505B scanner. Gene expression data from the scanned images were processed in the Agilent Feature Extraction software version 9.5.3.1 (Santa Clara, CA, USA).
3
Microarray analysis of ZnO nanoparticle effects
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Double-stranded cDNA was synthesized using the total RNA (250 ng) from individual mice (n ¼ 5 per experimental or control group) and Universal Mouse Reference total RNA (UMRR) (Agilent Technologies, Mississauga, ON, Canada). Cyaninelabelled cRNAs were synthesized from the cDNA using Quick Amp Labelling Kit (Agilent Technologies, Mississauga, ON, Canada). cRNAs from control and ZnO nanoparticle-treated samples were labeled with Cyanine 5-CTP, and reference cRNAs were labeled with Cyanine 3-CTP using a T7 RNA polymerase in vitro transcription kit (Agilent Technologies, Mississauga, ON, Canada) and purified using RNeasy Mini kits (Qiagen, Mississauga, ON, Canada). An equimolar amount of reference cRNA was mixed with each experimental cRNA sample and was hybridized to Agilent mouse 8 Â 60 k oligonucleotide microarrays (Agilent Technologies Inc., Mississauga, ON, Canada) for 17 h in a hybridization chamber at 65 C with a rotation speed of 10 rpm. Following hybridization, arrays were scanned on an Agilent G2505B scanner according to manufacturer's protocols. Gene expression data from the scanned images were extracted using Agilent Feature Extraction software version 9.5.3.1.
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