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30 minutes easy RNA extraction for RT-PCR! Kaneka Easy RNA Extraction Kit (for RT-PCR)

Date：May 23 2025Web Page No：520078

Quick and easy to prepare RNA for RT-PCR!

Kaneka Easy RNA Extraction Kit for RT-PCR can extract RNA for RT-PCR from blood and cultured cells by adding the reagents and just heating the samples. As the kit does not require irritated aqueous layer separation by using organic solvents, you can prepare RNA with quick and easy processing. This kit includes sufficient reagents for 100 preps.

Appearance of Easy RNA Extraction Kit

Features

  • Sample type: 103-106 cells, 0.5-2.0 μl blood (Heparin or EDTA treated)
  • Size: 100 preps
  • Operation time: Around 30 minutes

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Examples of Use

Results of PCR/RT-PCR using RNA extracted from cultured cells

Results of RNA extraction from cultured cells and RT-PCR detection

Adherent cells (HEK293T) and floating cells (Jurkat) were prepared to have 105 cells, respectively, and then RNA was extracted from the cells by using this kit. We used the extracts as RNA template and performed RT-PCR targeting ACTB mRNA. We analyzed the obtained RT-PCR products by electrophoresis and detected amplified fragments originated from the targeted mRNA (Figure 1, and 2). We also used the same extracts as template RNA and performed PCR in order to confirm that the extracts did not contain any genomic DNA from the original cells by electrophoresis. The result showed that any amplified fragments were not detected.


Results of PCR/RT-PCR using RNA extracted from mouse blood

Results of RNA extraction from mouse blood at RT-PCR detection

RNA was extracted from mouse blood with heparin as an anticoagulant by using this kit. We then used the extract as a template RNA and performed RT-PCR targeting H3f3a mRNA. We analyzed the obtained RT-PCR product by electrophoresis and detected the amplified fragment originated from the targeted mRNA (Figure 3). We also used the same extract as a template RNA and performed PCR in order to confirm that the extract did not contain any genomic DNA from the mouse blood by electrophoresis. The result showed that any amplified fragments were not detected.


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Protocol Overview

Image of Protocol overview

■Cultured cells (Click here!)
  1. Transfer the cell culture medium to a centrifuge tube. Adjust the number of cells to 103 ~ 106 per tube.
  2. Centrifuge the tube at 1,000 rpm for 3 minutes. Gently remove the medium components while not peeling off the collected cell pellets.
  3. Add 100 μl of PBS (-) to the cell pellet and resuspend the cells by pipetting. Centrifuge the tube at 1,000 rpm for 3 minutes, then remove the supernatant and collect the cells at the bottom of the tube.
  4. Add 20 μl of Reagent A to the centrifuge tube containing the cells and pipette well to resuspend the cells in the reagent. Transfer the cell suspension to a new PCR tube.
  5. Incubate the PCR tube containing the cell suspension in a heat block or other devices, at 75 °C for 5 minutes.
  6. After cooling down the PCR tube to room temperature, add 1 μl of DNase I and mix well by pipetting. Incubate the tube at 42 °C for 10 minutes and then at 75 °C for 5 minutes in a heat block or other devices.
  7. Now the prepared extract is ready for a template of RT-PCR system. Add the extract to the reaction system according to the protocol of the RT-PCR kit you use. For example, 1 to 2 μl of the extract *1 is used for the reverse transcription reaction (in the case of a 25 μl reaction system) *2.
■Blood samples (Click here!)
  1. Add 20 μl of Reagent A to the PCR tube.
  2. Add 0.5 to 2.0 µl of fresh blood containing an anticoagulant such as heparin *3 to the above PCR tube and mix well by pipetting.
  3. Incubate the above PCR tube in a heat block at 75 °C for 5 minutes.
  4. After cooling down the PCR tube to room temperature, add 1 µl of DNase I and mix by pipetting. Incubate at 42 °C for 10 minutes and then at 75 °C for 5 minutes in a heat block or other devices.
  5. Now the prepared extract is ready for a template of RT-PCR system. Add the extract to the reaction system according to the protocol of the RT-PCR kit you use. For example, 1 to 2 μl of the extract *1 is used for the reverse transcription reaction (in the case of a 25 μl reaction system) *2.

*1 Some precipitates may be seen in the incubated solution. It is not required to remove the precipitate. But in the case of removing the precipitate, centrifuge the PCR tube at 5,000 rpm for 5 minutes at 4 °C to collect the supernatant.
*2 It is recommended to use the extract as soon as possible after the procedure is completed. When using the extract, keep it on ice. In the case of storing, keep it at -20 °C or below.
*3 It has been confirmed that the kit is compatible for blood samples treated with heparin or EDTA as an anticoagulant.


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Kit Components

  • Reagent A
  • Deoxyribonuclease I (DNase I)

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Product Information

[Date : July 15 2025 00:07]

Detail Product Name Product Code Supplier Size Price
Kaneka Easy RNA Extraction Kit (for RT-PCR)
DatasheetThis may not be the latest data sheet. 		KN-T110004 KNK 1 kit $446

Description This kit can extract RNA for RT-PCR from blood and cultured cells for 30 minutes. No need to perform aqueous layer separation by using organic solvents. Sample : Blood, Cultured Cell, Time : -30 minutes Size : 100 preps
Storage -20°C CAS
Link

[Date : July 15 2025 00:07]

Kaneka Easy RNA Extraction Kit (for RT-PCR)

DatasheetThis may not be the latest data sheet.


  • Product Code: KN-T110004
  • Supplier: KNK
  • Size: 1kit
  • Price: $446

Description This kit can extract RNA for RT-PCR from blood and cultured cells for 30 minutes. No need to perform aqueous layer separation by using organic solvents. Sample : Blood, Cultured Cell, Time : -30 minutes Size : 100 preps
Storage -20°C CAS
Link

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CONTACT

export@funakoshi.co.jp

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