Prior to experimentation, HCT116-OxR cells and HT29-OxR cells were cultured 1 week without oxaliplatin. Total RNA from HCT116, HCT116-OxR, HT29 and HT29-OxR cell lines was isolated with Trizol reagent (Invitrogen, Carlsbad, CA) and miRNA fraction was further purified using a mirVana™ miRNA isolation kit (Ambion, Austin, TX). The isolated miRNAs was labeled with Hy3 using the miRCURY™ Array Labelling kit (Exiqon, Vedbaek, Denmark) and hybridized on a miRCURYTM LNA microRNA Array (v 8.0, Exiqon) as described [37 (link)]. Microarray images were acquired using a Genepix 4000B scanner (Axon Instruments, Union City, CA) and processed and analyzed with Genepix Pro 6.0 software (Axon Instruments) and Excel.
Mircury array labelling kit
Manufactured by Qiagen
Sourced in Denmark
The miRCURY™ Array Labelling Kit is a product designed for the labelling of microRNA (miRNA) samples prior to their analysis on miRNA microarray platforms. The kit provides the necessary reagents and protocols for efficient labelling of miRNA samples, enabling accurate and reliable quantification of miRNA expression levels.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (3 mentions)
Genepix pro 6, by Molecular Devices (3 mentions)
Mirvana mirna isolation kit, by Thermo Fisher Scientific (2 mentions)
Genepix 4000b scanner, by Molecular Devices (2 mentions)
Mircury lna microrna array, by Qiagen (1 mentions)
Axon genepix 4000b microarray scanner, by Molecular Devices (1 mentions)
Mirneasy mini kit, by Qiagen (1 mentions)
Mircury lna array, by Agilent Technologies (1 mentions)
Genepix 4000b laser scanner, by Molecular Devices (1 mentions)
Genepix 4000b array scanner, by Molecular Devices (1 mentions)
Mircury lna array, by Qiagen (1 mentions)
5 protocols using mircury array labelling kit
1
Microarray Analysis of miRNA Expression in Oxaliplatin-Resistant Cells
2
Nasal Mucosa microRNA Profiling in Allergic Rhinitis
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Preparation of nasal mucosal specimens. Nasal mucosal samples were obtained from inferior turbinate sections from 25 patients with perennial AR and 25 patients with nonallergic rhinitis (NAR control group) according to the method of Ruocco et al (18) . Diagnosed based on the case history, nasal endoscopy, allergen skin-prick tests, and specific IgE assays. No patient had received oral steroids or other medications for 4 weeks prior to study recruitment. The presemt study was approved by the Institutional Review Board at Hangzhou First People's Hospital, Nanjing Medical University. All participants provided written informed consent. microRNA expression profiling. Total RNA was extracted from nasal mucus from AR patients and NAR controls using the miRNeasy mini kit (Qiagen, West Sussex, UK) according to manufacturer's instructions. Samples were labeled with Hy3 using the miRCURY™ Array Labelling kit (Exiqon, Inc., Woburn, MA, USA) and hybridized to the miRCURY LNA™ Array (v.16.0; Agilent Technologies, Inc., Santa Clara, CA, USA). After washing, the chips were scanned with the Axon GenePix 4000B Microarray Scanner (Molecular Devices, LLC, Sunnyvale, CA, USA). The procedure and images process method as described previously (19) . The miRNA expressions of all differentially expressed samples were clearly displayed by a hierarchical clustering heat map.
3
miRNA Microarray Analysis of Lentiviral Infection
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miRNA microarray analysis was performed as previously described (30 (link),61 (link)). Total RNA from EA.hy926 cells either Mock-infected or infected with K1 or Nef lentivirus alone, or co-infected with both K1 and Nef lentiviruses for 72 h were isolated with Trizol reagent (Invitrogen). The miRNA fraction was further purified using a mirVanaTM miRNA isolation kit (Ambion, Austin, TX, USA). The isolated miRNAs from the four parallel infected cells were then labeled with Hy3 using the miRCURYTM Array Labelling Kit (Exiqon, Vedbaek, Denmark) and hybridized individually on miRCURYTM LNA microRNA Arrays (v 8.0; Exiqon). Microarray images were acquired using a Genepix 4000B scanner (Axon Instruments, Union City, CA, USA), processed and analyzed with Genepix Pro 6.0 software (Axon Instruments). EA.hy926 cells transduced with Mock were used as the control.
4
miRNA Profiling of Colon Cancer Co-culture
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Total RNA, including miRNA, was isolated using Trizol reagent (Invitrogen Life Technologies, Carlsbad, CA, USA) according to the manufacturer’s instructions from HLECs co-cultured with colon cancer cell lines.
The isolated miRNAs were then labeled with Hy3TM using the miRCURYTM Array Labelling kit (Exiqon, Vedbaek, Denmark) and then hybridized on miRCURYTM LNA microRNA Array 16.0 edition (Exiqon, Vedbaek, Denmark), as previously described [13 (link)]. Hybridization images were collected using a GenePix 4000B laser scanner (Molecular Devices, Sunnyvale, CA, USA). Images were quantified using GenePix Pro 6.0 (Axon Instruments, Sunnyvale, CA, USA). Raw data were further processed in Microsoft Excel.
The isolated miRNAs were then labeled with Hy3TM using the miRCURYTM Array Labelling kit (Exiqon, Vedbaek, Denmark) and then hybridized on miRCURYTM LNA microRNA Array 16.0 edition (Exiqon, Vedbaek, Denmark), as previously described [13 (link)]. Hybridization images were collected using a GenePix 4000B laser scanner (Molecular Devices, Sunnyvale, CA, USA). Images were quantified using GenePix Pro 6.0 (Axon Instruments, Sunnyvale, CA, USA). Raw data were further processed in Microsoft Excel.
5
Profiling Ascending Colon miRNA Expressions
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Shanghai KangChen Bio-tech measured the miRNA expression profiles of the ascending colon mucosa samples using the miRCURY TM LNA Array (v.16.0, Exiqon, Vedbaek, Denmark). The miRNA hybridization method was based on the procedures provided with the miRCURY LNA. RNA samples were labelled with Hy3 fluorescence using the miRCURY TM array labelling kit (Exiqon, Vedbaek, Denmark). Fluorescence-labelled miRNAs were identified after hybridization by sequencing. The hybridized miRNAs were washed using the buffer kit (Exiqon, Vedbaek, Denmark), dried, and scanned using a GenePix 4000B array scanner (Molecular Devices, Sunnyvale, CA, USA).
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