Thermo Scientific KingFisher Pure RNA Blood Kit, Инструкция по эксплуатации

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10 | Thermo Scientific KingFisher Pure RNA Blood Kit Thermo Fisher Scientific
Chapter 2 | Product Description
Thermo Scientific™ KingFisher™ instrument and the KingFisher Pure RNA
Blood Kit for any particular use, as the performance of the kits has not been
validated for any specific organism or downstream application.
Principle and Procedure
The KingFisher Pure RNA Blood Kit uses magnetic-particle technology for
RNA purification. The Thermo Scientific™ KingFisher™ technology combines
the speed and efficiency of RNA purification with easy handling of magnetic
particles. The purification process requires no phenol/chloroform extraction
and needs very little hands-on time.
The first step of the protocol lyses the cells, after which the NA (nucleic acid)
can bind to the surface of the KingFisher Magnetic Beads in the presence of a
chaotropic salt. Copurified DNA is removed during a DNase I treatment step.
The following effective wash steps dispose of proteins, cell debris, and any
residual contaminants, while the RNA bound to the KingFisher Magnetic
Beads is transferred through the wash steps.
Two different Wash Buffers are used, followed by an air drying step. High-
quality RNA is eluted into nuclease-free water, and is ready for subsequent
downstream processes.
Kit Specifications
The KingFisher Pure RNA Blood Kit is designed for rapid automated
preparation of highly pure total RNA from up to 200 µl of blood samples
using KingFisher magnetic particle processors.
If a dispense step requiring the addition of Rebinding Buffer is excluded, the
approximate processing time is 45 min for the purification of 96 samples
on the KingFisher Flex and 12 samples on the KingFisher Duo. The obtained
RNA can be used directly in various downstream applications.
Fresh or stored blood samples can be used. Suitable sample storage is
essential for obtaining a high yield and good quality total RNA.
Typically 1.5-4 µg of total RNA can be purified from a 200 µl volume of
blood sample with an A
260
/A
280
ratio of ≥ 1.8–2.1. The yields of acquired
purified RNA depend on the sample type, and the method of sample
collection, and storage. Refer to
Appendix B for typical total RNA yields from
various sources.