EXTRAClean Plasma/Serum RNA Purification Kit
EXTRAClean Plasma/Serum RNA Purification Kit
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Supporting Data
| Kit Specifications | |
| Sample Type | Plasma/Serum |
| Anti-Coagulant (for Plasma)† | EDTA or Citrate |
| Sample Volume Range | 50 to 200 μL |
| Minimum Elution Volume | 10 μL |
| Maximum Elution Volume | 25 μL |
| Time to Complete 10 Purifications | 15 – 20 minutes |
| Size of RNA Purified | All sizes, including miRNA and small RNA (<200 nt) |
| Average Yields¥ | Variable depending on specimen |
† This kit is suitable for the isolation of RNA from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR
¥ Please check page 5 for Average Plasma/Serum Yields and Common RNA Quantification Methods
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2
years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at
60°C if any salt precipitation is observed.
| Component | Cat. 73700 (50 preps) | Cat. 73710 (20 preps) | Cat. 73720 (10 preps) |
|---|---|---|---|
| Lysis Buffer A | 2 x 20 mL | 100 mL | 1 x 130 mL 1 x 30 mL |
| Wash Solution A | 18 mL* | 1 x 38 mL* 1 x 18 mL* |
38 mL* |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Elution Buffer F | - | - | 15 mL |
| EXTRACLean Micro Spin Columns | 50 | - | - |
| EXTRACLean Mini Spin Columns | - | 20 | 10 |
| EXTRACLean Midi Spin Columns | - | 20 | - |
| EXTRACLean Maxi Spin Columns | - | - | 10 |
| Collection Tubes | - | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 |
* For the preparation of working solutions, please see important Notes (Notes prior to use)
Documentation
FAQs
Mini, Midi, Maxi
A fixed speed centrifuge can be used, however reduced yields may be observed.
All centrifugation steps are performed at room temperature. Centrifugation at 4°C will not adversely affect kit performance.
Adding more or less than the specified volumes may reduce both the quality and the quantity of the purified RNA. Eluting your RNA in high volumes will increase the yield but will lower the concentration. Eluting in small volumes will increase the concentration but will lower the overall yield.
Your purified RNA will be contaminated with the Wash Solution A. This may reduce the quality of your purified RNA and will interfere with your downstream applications
Yes, but it is recommended that the 2nd elution be in a smaller volume (50% of 1st Elution). It is also recommended to perform the 2nd elution into a separate elution tube to avoid diluting the 1st elution
Plasma/Serum samples contain very little RNA. This varies from individual to individual based on numerous variables. In order to increase the yield, the amount of Plasma/Serum input could be increased.
Most of the Free-Circulating Plasma/Serum RNA is short RNA fragments. The A260:280 ratio is normally between 1 – 1.6. This low A260:280 ratio will not affect any downstream application.
If a different Elution Buffer was used other than the one provided in the kit, the buffer should be checked for any components that may interfere with the application. Common components that are known to interfere are high salts (including EDTA), detergents and other denaturants. Check the compatibility of your elution buffer with the intended use.
You may need to do a DNase treatment to your isolated Plasma/Serum miRNA. It is recommended to use Norgen’s RNase-Free DNase I Kit (Cat# 25710). Also please refer to the protocol for optional on-column DNA removal outlined in the protocols.