Total RNA was isolated from tissue blocks of corpus callosum/hippocampus using the Trizol Technique (Life Technologies, Invitrogen) and RNeasy Mini Kit 250 (Qiagen). Reverse transcription was performed using SuperScript II Reverse Transcriptase and dNTP set (Invitrogen), Random primers and RNasin (Promega) and followed by qPCR using the TaqMan Gene Expression Assays: PCR Master Mix, predesigned Taqman primers for MBP (Inventoried) and endogenous control (GAPDH), Thermo-Fast 96, and ABgene plates (Applied Biosystems, Thermofisher). Expression of genes was analyzed with the 7300 Systems SDS Software (Applied Biosystem) and MBP expression was normalized to reference gene GAPDH, whose expression was stable in different groups and in two independent experiments.
Random primers and rnasin
Manufactured by Promega
Random primers are short, synthetic DNA sequences that can bind to and amplify multiple regions of a nucleic acid sample. RNasin is an RNase inhibitor that helps protect RNA samples from degradation during experimental procedures.
Automatically generated - may contain errors
2 protocols using random primers and rnasin
1
Quantifying Myelination Markers in Tissues
2
Quantification of Oligodendrocyte Markers
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Total RNA was extracted from blocks of corpus callosum/hippocampus of 1‐week‐treated animals or controls, using the Trizol Technique (Life technologies, Invitrogen) and RNeasy mini‐kits 250 (Quiagen). Reverse transcription was performed using Superscript II reverse transcriptase and dNTP set (Invitrogen), Random primers and RNasin (Promega) and followed by qPCR using the TaqMan Gene Expression Assays: PCR master Mix, predesigned Taqman primers for Myt1, Olig2 and Sox17, and endogenous control (GAPDH), Thermo‐Fast 96 and ABgene plates (Applied Biosystems, Thermofisher). Expression of genes was analyzed with the 7300 Systems SDS Software (Applied Biosystem) and the expression of Myt1, Olig2, and Sox17 normalized to reference gene GAPDH whose expression was stable in different groups.
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