The RNA Clean & Concentrator-5 kit is an RNA clean-up kit that provides a simple and reliable method for the rapid preparation of high-quality, RT-PCR-ready, DNA-free (R1013, R1014) RNA. This simple procedure is based on the use of a unique single-buffer system and Zymo-Spin column technology that allows for selective recovery of total RNA (> 17 nt), large RNAs (> 200 nt), and/or small RNAs (17-200 nt). The procedure is easy: Add binding buffer and ethanol to your sample, then bind, wash and elute ultra-pure RNA. The RNA can be eluted from the Zymo-Spin IC Column in as little as ≥ 6 µl of RNase-free water. The highly-concentrated, purified RNA is suitable for all subsequent analyses and molecular manipulations.
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| Equipment | Microcentrifuge |
|---|---|
| Format | Spin-Column |
| Purity | RNA is ready for Next-Gen sequencing, RT-PCR, microarray, hybridization, etc. A260/A280, A260/A230: > 1.8. |
| Sample Source | DNase I treated RNA, in vitro transcription products, the aqueous phase following TRIzol/chloroform or similar extraction |
| Size Range | Total RNA ≥ 17 nt |
| Yield | 10 µg RNA (binding capacity), ≥ 6 µl (elution volume) |
Q1: Can the RCC kit be used to purify DNA (cDNA)?
Yes. The kit efficiently recovers all types of nucleic acids.
Q2: Should carrier RNA be added to the sample prior to purification?
Carrier RNA can be added with no detrimental effects. The RCC is highly efficient without the need for carrier RNA, and no significant improvement in recovery has been observed with carrier RNA.
Q3: Will this kit remove fluorescent dyes, radiolabeled dNTP’s and/or Biotin?
Yes, the kit efficiently removes unincorporated fluorescent dyes, radiolabeled dNTP’s and Biotin.
Q4: Are samples containing high concentration of detergents, salts, formamide or sucrose compatible with the RCC buffer system?
Product Tolerance Reference: – ≤5% Triton X-100 – ≤5% Tween-20 – ≤5% Sarkosyl – ≤0.1% SDS – ≤90% formamide – Sucrose samples should be diluted/titrated down 10- to 100- fold.
Q5: Can other DNase I enzymes or sets (e.g. Qiagen DNase I) be used?
Yes, follow respective protocol for on-column DNase treatment. If the DNase does not have a protocol, proceed with in-tube DNase treatment post clean-up, then re-purify.
Q6: Is DNase I treatment necessary?
If the downstream application requires DNA-free RNA, we would recommend performing in-column or in-tube DNase I treatment.
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