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[DESCRIPTION]
<div class="column">The steps for preparing the lysate are different depending on the starting material. Please ensure you follow the correct procedure for your starting material (see the section <a href="https://www.protocols.io/view/genomic-dna-removal-and-total-rna-purification-fro-f48bqzw?guidelines" target="_blank">Disrupting and Homogenizing Starting Materials</a> and the different lysate preparation protocols <a href="https://www.protocols.io/view/isolate-ii-biofluids-rna-kit-f5kbq4w" target="_blank">here</a>). The subsequent steps detailed in <a href="https://www.protocols.io/view/genomic-dna-removal-and-total-rna-purification-fro-f48bqzw" target="_blank">Genomic DNA removal and total RNA purification</a> are the same in all cases.</div>
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<p>This protocol describes the isolation of viral RNA from small amounts of tissue (up to 20 mg for most types of tissue).</p>

[BEFORE_START]
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<p>Ensure Lysis Buffer RX is prepared (see guidelines).</p>
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<p>Ensure that all solutions are at room temperature before use.</p>
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<p>Two types of spin columns are provided with this kit: the <strong>Genomic DNA Removal Column</strong></p>
<p>(blue ring) and the <strong>RNA Column</strong> (black ring). Ensure the correct column is used for each</p>
<p>step of the procedure.</p>
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<p>All centrifugation steps are carried out in a benchtop microcentrifuge at 14,000 x g except</p>
<p>where noted. Perform all centrifugation steps at room temperature.</p>
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<p>It is important to work quickly during this procedure.</p>
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[GUIDELINES]
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<li>This protocol describes the isolation of viral RNA from small amounts of tissue (up to 20 mg for most types of tissue). Refer to Table 1 as a guideline for maximum tissue input amounts. If your tissue of interest is not included in the table, we recommend starting with an input of no more than 10 mg.</li>
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<p><strong>TABLE 1: RECOMMENDED INPUT OF DIFFERENT TISSUES</strong></p>
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<p><strong>Tissue</strong></p>
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<p><strong>Max. Input Amount</strong></p>
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<p>Brain</p>
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<p>25 mg</p>
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<p>Heart</p>
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<p>5 mg</p>
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<p>Kidney, Liver, Lung, Spleen</p>
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<p>20 mg</p>
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<p>Other tissues</p>
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<p>10 mg</p>
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<p>RNA in mammalian tissue is not protected from RNases after harvesting until the tissue is disrupted and homogenized. Therefore, it is important that the procedure is carried out as quickly as possible.</p>
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<p>Fresh or frozen tissues may be used for the procedure. Tissues should be flash-frozen in liquid nitrogen and transferred immediately to a -80°C freezer for long-term storage. Tissues may be stored at -80°C for several months. Do not allow frozen tissues to thaw prior to grinding with the mortar and pestle in order to avoid compromising the integrity of the RNA.</p>
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<p>Please review the Guidelines under <a href="https://www.protocols.io/view/genomic-dna-removal-and-total-rna-purification-fro-f48bqzw?guidelines" target="_blank">Genomic DNA removal and total RNA purification from all types of lysate</a> for other important details.</p>
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