Real-time RT-PCR was performed using total RNA isolated on RNeasy Quick spin columns (QIAGEN, CA). One μg of total RNA was used to perform a reverse transcriptase-polymerase chain reaction (RT-PCR) using iScript supermix (Biorad, Hercules, CA, USA). The sequence of PCR primers used are: Vav1, 5′-CAACCTGCGTGAGGTCAAC-3′ and 5′-ACCTTGCCAAAATCCTGCACA-3′; VEGF, 5′-TGTACCTCCACCATGCCAAGT-3′ and, 5′-CGCTGGTAGACGTCCATGAA-3′; PDK1, 5′-ACCAGGACAGCCAATACAAG-3′, and 5′-CCTCGGTCACTCATCTTCAC-3′; Glut1, 5′-ACGCTCTGATCCCTCTCAGT-3′ and 5′-GCAGTACACACCGATGATGAAG-3′; EPO, 5′-ACCAACATTGCTTGTGCCAC-3′ and 5′-TCTGAATGCTTCCTGCTCTGG-3′. Values are expressed as fold increases relative to the reference sample (untreated control) and analyzed with CFX manager (Biorad). All primers were purchased from Sigma.
Rneasy quick spin columns
Manufactured by Qiagen
Sourced in United States
The RNeasy Quick spin columns are a core component of the RNeasy kit, designed for the rapid and efficient purification of high-quality RNA from a variety of sample types. The columns utilize a silica-based membrane to selectively bind RNA, allowing for effective removal of contaminants and impurities.
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Lab products found in correlation
2 protocols using rneasy quick spin columns
1
Real-time RT-PCR for Gene Expression
2
qRT-PCR Analysis of Gene Expression
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qRT-PCR was performed using total RNA isolated on RNeasy Quick spin columns (QIAGEN, Valencia CA, USA). One μg of total RNA was used to perform RT-PCR using iScript supermix (Biorad, Hercules, CA, USA). The sequence of PCR primers used is NGP, 5′-AGACCTTTGTATTGGTGGTGGC-3′ for sense and 5′-GGTTGTATGCCTCTATGGCTCTA-3′ for reverse; β-actin, 5′-CTGTCCCTGTATGCCTCTG-3′ for sense and 5′-ATGTCACGCACGATTTCC-3′ for reverse. Values are expressed as fold increase relative to β-actin and analyzed with CFX Manager (Biorad). All primers were purchased from Sigma-Aldrich.
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