These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples 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. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.
Background
Plasma/Serum cell-free circulating RNA or exosomal RNA has the potential to provide biomarkers for certain cancers and disease states. Exosomes are 40 – 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses they can be efficiently recovered from biological fluids, such as plasma or serum.
Plasma/Serum RNA Purification Mini Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 µL to 200 µL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 µL to 25 µL.
Plasma/Serum RNA Purification Midi Kit
This utilizes a two column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 µL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.
Plasma/Serum RNA Purification Maxi Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.
Isolate RNA after Purifying EVs and Exosomes
For Ultracentrifugation, Exoquick, and Filtration
| Cat. # | Name | Elution Volume | Plasma/Serum | Urine | Cell-Culture Media |
|---|---|---|---|---|---|
| 55000 | Plasma/Serum RNA Purification Mini Kit | 10 – 25 µL | 50 µL – 1 mL | 250 µL – 1 mL | 5 – 10 mL |
| 35300 | Total RNA Purification Micro Kit | 20 – 50 µL | 1 – 4 mL | 2 – 10 mL | 10 – 20 mL |
| 17200 | Total RNA Purification Kit | 50 – 100 µL | 4 – 10 mL | 11 – 30 mL | 20 – 35 mL |
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| Kit Specifications | |
| Minimum Plasma/Serum Input | 2 mL |
| Maximum Plasma/Serum Input | 5 mL |
| Size of DNA Purified | All sizes, including miRNA and small RNA (<200 nt) |
| Elution Volume | 50-100 μL |
| Time to Complete 10 Purifications | 35-40 minutes |
| Average Yields* | Variable depending on specimen |
*Please check page 7 of the Product Insert for Average Plasma/Serum Yields and Common RNA Quantification Methods.
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.
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.
| Kit Components | Cat. 55000 (50 preps) | Cat. 56100 (20 preps) | Cat. 56200 (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 | 15 mL |
| Micro Spin Columns | 50 | – | – |
| Mini Spin Columns | – | 20 | 10 |
| Midi Spin Columns | – | 20 | – |
| Maxi Spin Columns | – | – | 10 |
| Collection Tubes | 50 | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 |
Usages:
For selective enrichment of Salmonella.
Principle:
Peptone provide carbon and nitrogen sources to meet the needs of bacterial growth; lactose are fermentable sugars; selenite, sodium hydrogen inhibit Gram-positive bacteria and gram-negative enterobacteria most non-Salmonella; phosphate-buffered agent; L- cystine as a reducing agent.
Formulation(per liter):
Peptone: 5g
Lactose :4g
Sodium selenite: 4g
Disodium hydrogen phosphate: 10g
L- cystine: 0.01g
Final pH 7.0 ± 0.2
How to use:
1. Weigh 23g of the product , adding 1 L of distilled or deionized water , heated to boiling stirring until completely dissolved, dispensing flask, cooled to room temperature .
2. Pipette 10mL of pre-enrichment sample broth or 25mL and transferred species in the liquid sample flask in a sterile environment.
3.Place into incubator, cultured at 36 ± 1 for 18-24h.
4. Observe the results.
Quality control:
Quality control strains were inoculated ,and cultured at 36 ± 1 for 18-24h ,results show as follows:
strain name strain code growth feature
Salmonella typhi CMCC (B) 50071 good red, cloudy
Salmonella typhimurium CMCC (B) 50115 good red, cloudy
Escherichia coli ATCC25922 — remain unchanged
Storage: Store in a dark, cool and dry place, tighten the cap immediately after use. Storage period of three years.
500g
