Easy Purification Of Pure RNA From The Smallest Samples

Easy Purification of Pure RNA from the Smallest Samples

• Isolate Total RNA from 1 Cell to 1 x 10^{4 Cells}
• Small 10 µl Elution Volume
• Ideal for Microarray Applications

The Absolutely RNA™ nanoprep kit easily purifies DNA-free total RNA from small cell samples and elutes in a volume of 10 µl, providing pure RNA at a concentration useful for a variety of applications such as microarray target generation and quantitative RT-PCR. This kit includes a simple and efficient DNA removal step directly on the spin column, making it ideal for the most
sensitive applications.

Smallest Elution Volume

The Absolutely RNA™ nanoprep kit yields high-quality total RNA in a small 10-µl elution volume, ideal for quantitative RT-PCR and the preparation of microarray fluorescent targets. The kit is specially designed to isolate total RNA from small amounts of starting material, down to a single cell. Previously, the consequence of purifying RNA from such small samples is that often, the RNA is too dilute resulting in the need for additional concentration steps and the potential for introducing unwanted contamination. The Absolutely RNA nanoprep kit is optimized for total RNA elution in a 10-µl volume, yielding total RNA in a useful concentration from even the smallest samples. The Absolutely RNA nanoprep kit is ideal for quantitative RT-PCR (QRT-PCR) and microarray applications where isolating total RNA from very small samples is common. In addition, the kit can easily be used to obtain pure total RNA from laser microdissection samples without additional steps or preparation.

Fast and Easy Absolutely RNA™ Nanoprep Method

(Figure 1)

Fast and Easy Method

The Absolutely RNA nanoprep kit is designed for fast and easy isolation from samples from 1 cell to 10⁴ cells (Figure 1). Isolation of total RNA is performed in a single nano-spin cup to minimize potential loss of RNA during the procedure. Following cell lysis, the solution is mixed with ethanol and applied to the silica-based fiber matrix within the nano-spin cup to bind the RNA. DNA contamination is removed by incubation with DNase I directly on the column. The RNA bound to the fiber matrix is washed to remove contaminants and total RNA is recovered in 10-µl final elution buffer.

Reliable RNA Purification Down to a Single Cell
QRT-PCR results using total RNA purified with the Absolutely RNA™ nanoprep kit isolated from 10⁴, 10³, 10², 10 and 1 HeLa cell. Duplicate reactions were run for each sample with human GAPDH molecular beacon and primers and 2 µl RNA template. Reactions were performed with the Mx4000™ multiplex quantitative PCR system. (Figure 2)

Smallest Samples

Quantitative real-time RT-PCR is a powerful tool in understanding gene expression. Often, the samples used for analysis are very small where it is critical to have an effective method of isolating pure RNA. To demonstrate, QRT-PCR was performed using total RNA purified from HeLa cells ranging from 104 cells to one cell (Figure 2). The Absolutely RNA nanoprep kit provides a reliable method for purifying DNA-free RNA from samples as small as a single cell.

Perfect for the Smallest Preps

The Absolutely RNA nanoprep kit contains all the necessary components for fast and easy DNA-free total RNA purification for extremely small samples. DNase I is provided lyophilized for convenient room-temperature storage, saving valuable freezer space. The small elution volume provides RNA in a useful concentration for the smallest of samples, making the Absolutely RNA nanoprep kit ideal for many sensitive applications.

Stratagene also has Absolutely RNA kits for larger samples. The Absolutely RNA™ Microprep kit isolates total RNA from 1 cell to 5 x 10⁵ cells in a 30-µl elution volume, and the Absolutely RNA™ RT-PCR Miniprep kit isolates total RNA from 10⁵ to 10⁷ cells or 5 to 40 mg of tissue in 50-µl elution volume.

Contributing Scientists
Stratagene: Scott Bashore and Gothami Padmabandu