BasicExoRNA™ Basic RNA extraction kit (Pre-isolated exosomes, 20 reactions)

Details for Product No. ABIN6240777,
RNA Extraction (REx)
Purpose RNA extraction Kit (from human biofluids and cell media)
Brand BasicExoRNA™
Sample Type Urine, Tissue Culture Medium, Cell Culture Cells, RNA Samples
Characteristics BasicExoRNA™ Kit (basic RNA extraction kit) allows total RNA (miRNA + mRNAs) extraction from pre-isolated exosomes pre-isolated via different methods including ultracentrifugation, chemical precipitation, immunocapture, size-chromatography etc). *RNA basic kit does not contain immunobeads for exosome isolation. Pre-isolated exosomes are lysed with an optimized lysis buffer and total RNA is purified using spin columns with a fast and user-friendly process. Eluted RNA can be used for downstream analyses or stored at -80 °C. Exosome standards for positive control are also included in the kit. All our kits guarantee high specificity for exosomal RNA and high yield of total RNA (including small RNAs) than similar products.
Samples: Pre-isolated exosomes from different samples (biofluids or cell culture media, either pellet after precipitation, frozen, or lyophilized).
Components Lysis buffer, RNA Washing buffer (to add Ethanol 96%), Elution buffer, Columns, Elution tubes, Lyophilized Exosome Standard
Material not included
  • Single-use and/or pipettes with disposable tips 2-100 μL
  • Pipettes 1 mL and 5 mL for reagent preparation
  • PBS
  • Disposable pipetting reservoirs
  • Ethanol 96 %
  • Chloroform or BCP (1-bromo-3-chloropropane)
  • Sample concentrator (urine and cell culture supernatant samples)

Further details regarding sample type: RNA extraction from Plasma, Serum, Urine, Cell culture media pre-isolated exosomes
Pre-isolated Exosomal RNA extraction

    BasicExoRNA™ Assay Protocol:
  1. RNA Extraction:
    • Lysis:
      1. Add 700 μL of Lysis buffer directly onto the exosome preparation.
      2. Resuspend by pipetting up and down until the lysate is clear.
      3. Incubate for 5 min at room temperature.
    • Extraction:
      1. Add 70 μL of 1-Bromo-3-chloropropane (BCP) or 140 μL of pure Chloroform.
      2. Shake for 30 sec.
      3. Incubate for 10 min at room temperature.
      4. Incubate for 1 min on ice and centrifuge at 12,000g at 4 °C for 10 min.
      5. NOTE: Incubation on ice prior to centrifugation helps in reducing DNA contamination, which tend to remain in the interphase.
      6. Transfer the top phase (aqueous) to a fresh tube.
      7. Add 2X of ethanol 96 % . Mix by gently inverting 4-5 times. If the top phase volume is 400 μL, add 800 μL of ethanol 96 %.
    • Purification:
      1. Transfer the half volume of the mixture into the spin column.
      2. Spin at 14,000g for 30 sec.
      3. Discard the flow-throw.
      4. Add the remaining volume into the same spin column.
      5. Spin at 14,000g for 30 sec. Discard the flow-throw.
      6. Wash the column with RNA Washing buffer. Add 400 μL of RNA Washing buffer in the column. Spin at 14,000g for 30 sec. Discard the flow-through. Perform the washing step twice more.
      7. Spin for 5 additional min at 14,000g to eliminate any ethanol residues from the column. Discard the flow-throw.
      8. Remove the tube and transfer the spin column into an elution tube.
      9. Elute the column with 15 μL of Elution buffer. Incubate for 5 min at room temperature. Spin for 2 min at 200 g and 1 min at 14,000g. Keep the flow-through. Eluted RNA is now ready for downstream analysis or for storage at -80 °C
Reagent Preparation
  • RNA Washing Buffer. Add into the bottle containing RNA Washing Buffer the volume of pure ethanol (96 % ) indicated on the bottle's label to get the final ethanol concentration of approximately 70 %.
  • Elution buffer and Lysis buffer are ready to use.
  • The RNase free column and elution tubes should be stored at room temperature.
  • All the opened buffers and diluted reagents including the bead washing buffer, RNA washing buffer, lysis buffer and the elution buffer should be stored at 4 °C.
Restrictions For Research Use only
Storage 4 °C
Storage Comment All the reagents are shipped and stored at +4°C for up to 24 months, if unopened. Briefly centrifuge small vials prior to opening. DO NOT FREEZE!
Expiry Date 24 months