RNA was isolated from 5000 to 10,000 sorted cells using the Arcturus PicoPure RNA Isolation Kit (Applied Biosystems, cat. no. 12204–01). After DNAse treatment (Qiagen, cat. no. 79254), complementary DNA (cDNA) was synthesized using qScript cDNA SuperMix (Quanta Biosciences, cat. no. 95048–100). RNA quality was confirmed with an Agilent Bioanalyzer 2100nusing Pico chip. Antral stomach was flash frozen in liquid nitrogen and RNA extracted using TRIzol (Invitrogen, cat. no. 15596026), loaded onto an RNeasy Mini Kit column (Qiagen, cat. no. 74106), and cDNA prepared using qScript cDNA SuperMix (Quanta Biosciences, cat. no. 95048–100). Quantitative PCR was performed using GoTaq qPCR Master Mix (Promega) in a Bio-Rad CFX96 system, and gene expression levels were determined by the ΔΔCt method with β-actin, GAPDH, or E-cadherin as controls. Primers are listed in the Table A1 .
Bioanalyzer 2100nusing pico chip
Manufactured by Agilent Technologies
The Bioanalyzer 2100n is a lab equipment product from Agilent Technologies. It utilizes Pico chips to perform automated electrophoretic analysis of DNA, RNA, and proteins.
Automatically generated - may contain errors
2 protocols using bioanalyzer 2100nusing pico chip
1
RNA Isolation and Gene Expression Analysis
2
RNA Isolation and Gene Expression Analysis
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RNA was isolated from 5000 to 10,000 sorted cells using the Arcturus PicoPure RNA Isolation Kit (Applied Biosystems, cat. no. 12204–01). After DNAse treatment (Qiagen, cat. no. 79254), complementary DNA (cDNA) was synthesized using qScript cDNA SuperMix (Quanta Biosciences, cat. no. 95048–100). RNA quality was confirmed with an Agilent Bioanalyzer 2100nusing Pico chip. Antral stomach was flash frozen in liquid nitrogen and RNA extracted using TRIzol (Invitrogen, cat. no. 15596026), loaded onto an RNeasy Mini Kit column (Qiagen, cat. no. 74106), and cDNA prepared using qScript cDNA SuperMix (Quanta Biosciences, cat. no. 95048–100). Quantitative PCR was performed using GoTaq qPCR Master Mix (Promega) in a Bio-Rad CFX96 system, and gene expression levels were determined by the ΔΔCt method with β-actin, GAPDH, or E-cadherin as controls. Primers are listed in the Table A1 .
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