Product/Service

Mini RNA Isolation Kit™

Source: Zymo Research
The Mini RNA Isolation Kit™ is an innovative product designed for easy, reliable, and rapid isolation of total RNA from small amounts of cells such as needle biopsy,
Highlights
  • Designed to isolate trace amounts of RNA.
    Simply and reliably isolates total RNA from 101-105 tissue culture cells.
  • Ideal for total RNA recovery for small sample amounts such as: needle biopsy, sorted cells, micro-dissected tissue, etc.
  • Total procedure in less than 1 hour.

General Description
The Mini RNA Isolation Kit™ is an innovative product designed for easy, reliable, and rapid isolation of total RNA from small amounts of cells such as needle biopsy, sorted cells, micro-dissected tissue, etc. The system combines our Zymo-Spin Column with a one-step RNA extraction and binding method. It makes ready-to-use RNA in less than 1 hour. The kit efficiently recovers high-quality total RNA for any subsequent analysis of gene expression. The Mini RNA Isolation Kit efficiently recovers total RNA from 1-105 cells. There is no phenol and chloroform used in this method.
Method Principle: As outlined in Figure 1, simply add RNA Extraction Buffer to the sample to extract RNA from cells. RNA is purified by Zymo-Spin Columns and pure RNA is eluted into small volumes, 8 ul, for convenient downstream manipulations.

Protocol
Before starting: Add 16 ml of 100% ethanol to the RNA Wash Buffer Concentrate.

    1. Cell Homogenization
    The kit is designed for efficient total RNA recovery from 10 to 104-105 cells. Further increasing the amount of cells over 106 will not improved the RNA recovery and may overload the system.
      A. Cell Sample: Collect cells by gentle centrifugation (2000 rpm on most centrifuges or about 400 g) for 5 minutes and remove the supernatant completely. Add 200 ul of RNA Extraction Buffer to the cell pellet. Lightly vortex to resuspend the pellet.
      B. Tissue Sample: Add 200 ul of RNA Extraction Buffer to the tissue sample. Grind the tissue using a homogenizer such as glass-teflon or Polytron devices in the RNA Extraction Buffer.

    2. Incubate the tube on ice for 20 minutes. Lightly vortex the tube briefly every 10 minutes.
    3. Add 1 volume (200 ml) of 95-100% Ethanol to the tube. Mix briefly. Incubate the tube on ice for 10 minutes.
    4. Transfer the mixture to a Zymo-Spin Column and place the column into a 2 ml Collection Tube.
    5. Spin the column and tube at full speed in a microcentrifuge for 1 minute.
    Empty the flow-through from the collection tube whenever necessary to prevent contamination of the column by the flow-through. The collection tube can hold 600 ul of liquid when the column is inserted.
    6. Add 200 ul of RNA Wash Buffer to the Zymo-Spin Column and centrifuge at full speed for 1 minute to wash.Add another 200 ul RNA Wash Buffer and repeat the microcentrifuge step as above.
    7. Transfer the column to a new 1.5 ml tube.
    8. Add 10 ul of RNA Elution Buffer directly to the membrane of Zymo-Spin Column. Wait for 2 minute. Spin briefly to elute RNA.

The eluted RNA now can be used immediately for any purpose or stored at -70°C for future use.

Comments

    1.RNA Storage
    It is recommended to add 1 unit of RNase inhibitor to the eluted RNA if the sample will not be used immediately. The RNA should be stored at -70°C for future use.
    2.DNA Contamination
    Generally, traces of fragmented DNA are present in the purified RNA. DNA-free RNA can be obtained by DNase I digestion.
    3. RNase contamination
    Use gloves. All the tubes and tips used should be RNase-free throughout the procedure.

Zymo Research, 625 West Katella Ave, Suite 32, Orange, CA 92867-4619. Tel: 714-288-9682; Fax: 714-288-9643.