Genomic DNA was extracted from frozen mononuclear cells of bone marrow samples by using the QIAamp DNA mini kit (Qiagen GmbH, Hilden, Germany) following the manufacturer’s instructions. The TRIzol Reagent (Life Technology, Carlsbad, CA) was used to extract RNA that was reversely transcribed to cDNA with the Superscript II RNase H2 reverse transcriptase kit (Invitrogen Corporation, Carlsbad, CA).
Superscript 2 rnase h2 reverse transcriptase kit
Manufactured by Thermo Fisher Scientific
Superscript II RNase H2 reverse transcriptase kit is a laboratory product designed for the reverse transcription of RNA into cDNA. The kit contains the necessary components, including the Superscript II RNase H2 reverse transcriptase enzyme, to facilitate this conversion process.
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2 protocols using superscript 2 rnase h2 reverse transcriptase kit
1
Genomic DNA and RNA Extraction
2
Quantitative Gene Expression Analysis
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Total RNA was extracted by using RNeasy Mini Kit (Qiagen) and underwent DNase treatment (Ambion Inc, Austin, TX). The quantity and quality of total RNA were assessed using a spectrophotometer (NanoDrop 1000; NanoDrop, Wilmington, DE). Reverse-transcription was performed using the Superscript II RNase H2 reverse transcriptase kit (Invitrogen). The relative abundance of transcripts was detected by quantitative (q) RT-PCR (KAPA SYBR FAST qPCR Master Mix; Stratagene, La Jolla, CA). The housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used to normalize the fluorescence level. The primers used for qRT-PCR are listed in Supplementary Table S2 .
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