Bile Salt Export Pump (BSEP) is a member of the ATP-binding cassette (ABC) transporters, which mediates the transport of bile acid, taurocholate and other cholate conjugates across the hepatocyte canalicular membrane into the canaliculus. BSEP is associated with progressive familial intrahepatic cholestasis type 2 (PFIC2) and benign recurrent intrahepatic cholestasis type 2 (BRIC2). PFIC2 caused by mutations in the BSEP gene increases the risk of hepatocellular carcinoma in early life.
This kit provides a rapid method for the isolation and purification of both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. The kit can be used to isolate all sizes of RNA from the cytoplasmic and nuclear RNA fractions, including all small RNA species without any requirement for phenol. Included in the kit are sufficient reagents to perform either 50 cytoplasmic RNA preparations or 25 cytoplasmic and 25 nuclear RNA preparations. Ten samples can be processed in approximately 45 minutes. This kit is also available in a 100 prep size.
Background
In certain circumstances it is desirable to be able to isolate fractionated RNA as opposed to total RNA. For example, it may be preferable to isolate only mature, cytoplasmic RNA for some studies on expression profiling. Alternatively it may be desirable to isolate nuclear RNA in order to investigate and study pre-processed (non-spliced) RNA. Furthermore, this kit can be used to isolate RNA for downstream applications where it is necessary to avoid DNA contamination, since the cytoplasmic fraction has been shown to be free of all traces of genomic DNA.
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| Kit Specifications | |
| Maximum Column Binding Capacity | Up to 50 µg RNA |
| Maximum Column Loading Volume | 650 µL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Time to Complete 10 Purifications | 45 minutes |
| RNA Yield HeLa (1 x 106) – Cytoplasmic RNA HeLa (1 x 106) – Nuclear RNA | 15 µg ≤ 3.5 µg |
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.
| Component | Cat. 21000 (50 preps) | Cat. 37400 (100 preps) |
|---|---|---|
| Lysis Buffer J | 20 mL | 2 x 20 mL |
| Buffer SK | 40 mL | 2 x 40 mL |
| Wash Solution A | 38 mL | 2 x 38 mL |
| Elution Buffer E | 6 mL | 2 x 6 mL |
| Mini Spin Columns | 50 | 100 |
| Collection Tubes | 50 | 100 |
| Elution Tubes (1.7 mL) | 50 | 100 |
| Product Insert | 1 | 1 |
The Flex caddy allows the Thermocycler GEN2 to be installed on the Opentrons Flex deck and sit below the deck, enabling below-deck cable routing and labware placed on top of modules to remain closer to the deck surface. The calibration adapter is used to calibrate the location of the Thermocycler. Caddies and calibration adapters are specific to the type of module.
Note: This item does not include the Thermocycler Module. The module is available with or without the Flex Caddy and Calibration tool: Thermocycler Module GEN2
The Flex caddy allows the Thermocycler GEN2 to be installed on the Opentrons Flex deck and sit below the deck, enabling below-deck cable routing and labware placed on top of modules to remain closer to the deck surface. The calibration adapter is used to calibrate the location of the Thermocycler. Caddies and calibration adapters are specific to the type of module.
Note: This item does not include the Thermocycler Module. The module is available with or without the Flex Caddy and Calibration tool: Thermocycler Module GEN2
