Total RNA was isolated using TRIzol (Ambion) from salivary glands (10–12 animals per sample is sufficient) vortexed at 4°C for 2 h or from S2 cell pellets vortexed for 30 min, extracted with ethanol precipitation, and subsequently purified with PureLink RNA Kit columns (Invitrogen). 1 μg of the extracted RNA was used for first-strand cDNA synthesis using a one-step RT-PCR kit (QIAGEN). To measure mRNA levels, real-time qPCRs were performed on resulting cDNA using gene-specific primers, as listed in Table S1 . Each RT-qPCR was repeated at least three times, the values were normalized to the Rp49 transcript. For X chromosome target genes where we compared both male and female expression, data were normalized to female controls. Error bars represented the standard error of the mean.
Purelink rna kit columns
Manufactured by Thermo Fisher Scientific
The PureLink RNA Kit columns are designed for the purification of total RNA from a variety of sample types, including cell lines, tissues, blood, and plants. The columns utilize a silica-based membrane to selectively bind RNA, allowing for the removal of contaminants and the elution of high-quality RNA.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using purelink rna kit columns
1
Quantifying Salivary Gland RNA Levels
2
Quantifying Salivary Gland RNA Levels
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Total RNA was isolated using TRIzol (Ambion) from salivary glands (10–12 animals per sample is sufficient) vortexed at 4°C for 2 h or from S2 cell pellets vortexed for 30 min, extracted with ethanol precipitation, and subsequently purified with PureLink RNA Kit columns (Invitrogen). 1 μg of the extracted RNA was used for first-strand cDNA synthesis using a one-step RT-PCR kit (QIAGEN). To measure mRNA levels, real-time qPCRs were performed on resulting cDNA using gene-specific primers, as listed in Table S1 . Each RT-qPCR was repeated at least three times, the values were normalized to the Rp49 transcript. For X chromosome target genes where we compared both male and female expression, data were normalized to female controls. Error bars represented the standard error of the mean.
3
RNA Extraction and qPCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total RNA was isolated using Trizol (Ambion) from salivary glands vortexed at 4°C for 2 h or from S2 cell pellets vortexed for 30 min, extracted with ethanol precipitation, and subsequently purified with PureLink RNA Kit columns (Invitrogen). 1 µg of the extracted RNA was used for first-strand cDNA synthesis using a one-step RT-PCR kit (Qiagen). To measure mRNA levels, real-time qPCRs were performed on resulting cDNA using gene-specific primers, as listed in Table S1.
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