This kit provides a rapid, single column method for the isolation and purification of total RNA (including miRNA) and proteins sequentially from a single sample of cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants. The total RNA and proteins are both column purified in under 25 minutes using a single column.
Purified RNA is of a high quality and yield, and is suitable for NGS, RT-qPCR and microarrays.
This kit eliminates DNA efficiently using a gDNA removal column.
Proteins are eluted in buffer and are ready for downstream applications such as Western Blots, Mass Spec and ELISA. The proteins will not require precipitation, resuspending of pellets, or any further cleaning.
This kit is ideal for researchers who are interested in studying the transcriptome and proteome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen’s RNA/Protein Purification Plus Kit is especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as biopsy materials or single foci from cell cultures, as it eliminates the need to fractionate the sample. Furthermore, analysis will be more reliable since the RNA and proteins are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications.
Protocol
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| Kit Specifications | |
| Maximum Column Binding Capacity | 50 μg for RNA |
| Maximum Column Binding Capacity | 200 μg for Protein |
| Maximum Column Loading Volume | 650 μL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Time to Complete 10 Purifications | 30 minutes |
| Average Yields*: HEK 293 Cells (1 x 106 cells) HEK 293 Cells (1 x 106 cells) Liver (15 mg) Liver (15 mg) | 10-15 μg RNA 70-100 μg protein 30-35 μg RNA 100-150 μg protein |
* Average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.
Storage Conditions and Product Stability
The Protein Loading Dye should be stored at -20°C after the addition of DL-Dithiothreitol (DTT). All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 48200 (50 preps) |
|---|---|
| Buffer SKP | 40 mL |
| Wash Solution A | 38 mL |
| Elution Solution A | 6 mL |
| Wash Solution C | 30 mL |
| Binding Buffer A | 8 mL |
| Elution Buffer C | 4 mL |
| Protein Neutralizer | 4 mL |
| Protein Loading Dye | 2 mL |
| gDNA Removal Columns | 50 |
| RNA/Protein Purification Columns | 50 |
| Collection Tubes | 150 |
| Elution Tubes (1.7 mL) | 100 |
| Product Insert | 1 |
Norgen’s FBS Exosome Depletion Kits provides a quick and easy protocol for the depletion of bovine exosomes from FBS prior to using it as a growth supplement in your culture medium. The FBS recovered from the depletion process is exosome-depleted and does not contain any quantifiable bovine miRNAs. Moreover, the exosome-depleted FBS will support the growth of your cells of interest similar to the non-depleted FBS. Norgen’s kits allow for the processing of different FBS volumes. The depletion is based on Norgen’s proprietary resin. These kits provide a clear advantage over other available kits in that they do not require ultracentrifugation, any special instrumentation, precipitation reagents or any protease treatments. More importantly, the depletion process is an inexpensive method for exosome depletion from FBS, as compared to the expensive current ready-to-use exosome-depleted media available on the market.
Background
Most culture medium used for the growth and propagation of cells in culture require the addition of fetal bovine serum (FBS) as a growth supplement to media. FBS is obtained from bovine (cow) serum, and therefore contains large quantities of bovine exosome vesicles. These exosomes may interfere with some types of studies, or may lead to unreliable results when studying the exosomes shed from your cells of interest in normal culture conditions. Therefore, the use of exosome-depleted FBS is highly recommended for many types of studies.
FBS Exosome Depletion Kit (Slurry Format)
For FBS volumes ranging from 140 mL to 280 mL.
FBS Exosome Depletion Kit (Column Format)
For FBS volumes ranging from 120 mL to 240 mL.
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| Kit Specifications | |
| Sample Type | Fetal Bovine Serum |
| Sample Volume Range | Up to 140 mL (FBS Exosome Depletion Kit I (Slurry Format) Up to 280 mL (FBS Exosome Depletion Kit II (Slurry Format) |
| Depletion | Deplete exosome-sized vesicle |
| Bovine miRNA | No detectable bovine miRNA |
| Time to Complete 6 Purifications | 40 minutes |
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
| Component | Cat. 61100 (6 preps) | Cat. 61400 (12 preps) | Cat. 61200 (6 preps) | Cat. 61300 (12 preps) |
|---|---|---|---|---|
| ExoC Buffer | 2 x 1.5 mL | 8 mL | 2 x 1.5 mL | 8 mL |
| Slurry E | 12.5 mL | 2 x 12.5 mL | – | – |
| Maxi Spin Column | – | – | 6 | 12 |
| Product Insert | 1 | 1 | 1 | 1 |
Human Cytomegalovirus (HCMV) or Human betaherpesvirus 5 (HHV-5) is a member of the Family Orthoherpesviridae. HCMV belongs to the betaherpesvirinae subfamily, while other herpesviruses fall into the subfamilies of Alphaherpesvirinae (including HSV 1 and 2 and varicella) or Gammaherpesvirinae (including Epstein-Barr virus). All herpes viruses share a characteristic ability to remain latent within the body over long periods of time. While CMV can be found in numerous body fluids including urine, saliva, breast milk, blood, tears, semen, and vaginal fluids, urine samples are generally used for congenital HCMV infection screening due to high viruria observed in infected infants.
CMV TaqMan PCR Kit, 100 reactions
CMV TaqMan PCR Probe/Primer Set and Controls, 100 reactions
For research use only and NOT intended for in vitro diagnostics.
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Storage Conditions and Product Stability
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
| Component | Cat. TM36350 (100 preps) | Cat. TM36310 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| CMV Primer & Probe Mix | 280 μL | 280 μL |
| CMV Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
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