Thermo Scientific KingFisher Pure RNA Blood Kit Instruction Manual
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Thermo Scientific KingFisher Pure RNA Blood Kit Instruction Manual

Thermo Scientific KingFisher Pure RNA Blood Kit Instruction Manual

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Thermo Scientific
KingFisher Pure RNA
Blood Kit
Revision 1.0
N14035 March 2013

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Summary of Contents for Thermo Scientific KingFisher Pure RNA Blood Kit

  • Page 1 Thermo Scientific KingFisher Pure RNA Blood Kit Revision 1.0 N14035 March 2013...
  • Page 2 Copyright © 2013 Thermo Fisher Scientific Inc. All rights reserved. PAXgene is a trademark of PreAnalytiX GmbH (a QIAGEN / BD Company). Tempus is a trademark of Life Technologies and its subsidiaries. Tween is a trademark of ICI Americas Inc. Virkon is a trademark of E.I. du Pont de Nemours and Company or its affiliates.

  • Page 3: Table Of Contents

    Ordering Information ....... . 33 NOTE: For more details on storing the kit reagents, refer to “Storage Conditions” page Thermo Fisher Scientific Thermo Scientific KingFisher Pure RNA Blood Kit...

  • Page 5: Kit Content

    30 ml * Addition of ethanol or isopropanol required. The KingFisher Pure RNA Blood Kit (Cat. No. 98020196) is intended for the purification of blood samples, using the Thermo Scientific™ KingFisher™ Flex with a 96 deep well head or the Thermo Scientific™ KingFisher™ Duo with a 12-pin head and a sample volume of 200 µl.

  • Page 6: Storage Conditions

    -20°C. The Proteinase K solution is stable at room temperature until the seal of the vial is broken. After being opened, the vial should be stored at -20°C. Store the Thermo Scientific™ KingFisher™ Magnetic Beads at +4°C. Other kit components can be stored at room temperature (15−25°C).
  • Page 7 Microtiter deep well 96 plate, sterile 50 pcs 97003530 KingFisher Duo Combi pack for Microtiter deep well 1 box 96 plate (tip combs, plates, and elution strips for 96 samples) Thermo Fisher Scientific Thermo Scientific KingFisher Pure RNA Blood Kit...

  • Page 9: Product Description

    3) Human buffy coat, blood leucocytes, erythrocytes, and bone marrow samples The RNA purified using the KingFisher Pure RNA Blood Kit is of high quality and free of proteins, nucleases, and other contaminants or inhibitors. It is,...

  • Page 10: Principle And Procedure

    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.

  • Page 11: Kingfisher Magnetic Particle Processors

    20 to 5000 µl. Each system consists of an instrument, specially designed plastic consumables, and the easy-to-use BindIt Software 3.2. The KingFisher Pure RNA Blood Kit is optimized and ready for use with the KingFisher Flex or KingFisher Duo.
  • Page 12 Heating block temperature from temperature +5°C above ambient room +10°C to +75°C, elution strip temperature to +115°C +4°C to +75°C at room temperature * See the details above on the Plates row. Thermo Scientific KingFisher Pure RNA Blood Kit Thermo Fisher Scientific...

  • Page 13: Safety Information

    Safety Information The following components of the KingFisher Pure RNA Blood Kit contain hazardous contents (Table 3-1). Always wear a laboratory coat, disposable gloves and goggles, and follow the safety instructions provided in the kit instruction manual. It is recommended that Good Laboratory Practice (GLP) is followed to guarantee reliable analyses.
  • Page 14 In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable protective clothing and gloves. This material and its container must be disposed of as hazardous waste. Thermo Scientific KingFisher Pure RNA Blood Kit Thermo Fisher Scientific...

  • Page 15: Storage Conditions And Preparation Of The Reagents

    Rebinding Buffer Add 100% isopropanol and 96−100% ethanol to the Wash Buffer 1, Wash Buffer 2, and Rebinding Buffer, as indicated below in Table 4-1 prior to the first use. Thermo Fisher Scientific Thermo Scientific KingFisher Pure RNA Blood Kit...

  • Page 16: Preparation Of The Dnase I Storage Solution

    -80°C for several months. Blood samples collected in PAXgene or Tempus tubes should be stored according to the manufacturer’s instructions. Thermo Scientific KingFisher Pure RNA Blood Kit Thermo Fisher Scientific...

  • Page 17: Protocols And Pipetting Instructions

    Thermo Scientific™ BindIt™ Software for KingFisher Instruments version 3.2 User Manual (Cat. No. N07974). BindIt Software protocols for the KingFisher Pure RNA Blood Kit can be found in BindIt Software 3.2 and at www.thermoscientific.com/kingfisher. Handling of KingFisher Magnetic Beads...

  • Page 18: Instructions For Kingfisher Flex With 96 Deep Well Plates

    KingFisher Pure RNA Blood Kit (Cat. No. 98020196) and the KingFisher Flex with Thermo Scientific™ Microtiter™ deep well 96 plates. When using the KingFisher Pure RNA Blood Kit for the first time, prepare the DNase I storage solution, Rebinding Buffer, Wash Buffer 1, and Wash Buffer 2.
  • Page 19 Chapter 5 Protocols and Pipetting Instructions 3. Place a Thermo Scientific™ KingFisher™ Flex 96 tip comb for deep well magnets on a Tip Plate (i.e. an empty KingFisher Flex 96 KF plate). 4. Start the PURE_RNABlood_Flex96 protocol using the KingFisher Flex 96 and load the plates as instructed on the KingFisher Flex 96 instrument display.

  • Page 20: Instructions For Kingfisher Duo With 12-Pin Magnet Head

    KingFisher Pure RNA Blood Kit (Cat. No. 98020196) and the KingFisher Duo with 12-pin magnet head. When using the KingFisher Pure RNA Blood Kit for the first time, prepare the DNase I storage solution, Rebinding Buffer, Wash Buffer 1, and Wash Buffer 2.
  • Page 21 KingFisher Duo elution strip Nuclease-free water 75 µl 4. Place a Thermo Scientific™ KingFisher™ Duo 12-tip comb into row B on the Blood RNA plate. 5. Start the PURE_RNABlood_Duo protocol using the KingFisher Duo and load the plate and elution strip.
  • Page 22 9. Place the plate back into the instrument and press OK. After the pause, the protocol will continue to completion. 10. After the run is completed, remove the plate and elution strip, and store the purified RNA. Thermo Scientific KingFisher Pure RNA Blood Kit Thermo Fisher Scientific...

  • Page 23: Instructions For Total Rna Purification From Paxgene Stabilized Blood Samples

    Buffer (supplemented with DTT) and 20 µl of Proteinase K. Mix the lysate and continue the purification as in “Instructions for KingFisher Flex with 96 Deep Well Plates” page 18 or as in “Instructions for KingFisher Duo with 12-pin Magnet Head” page Thermo Fisher Scientific Thermo Scientific KingFisher Pure RNA Blood Kit...

  • Page 24: Instructions For Total Rna Purification From Tempus Stabilized Blood Samples

    ). One unit at 260 nm corresponds to 40 μg of RNA per ml. The ratio between the A indicates the purity of the RNA. The value for RNA should be ≥ 1.8–2.1. Thermo Scientific KingFisher Pure RNA Blood Kit Thermo Fisher Scientific...
  • Page 25 ) x dilution factor • Total amount of RNA isolated = concentration x volume of sample in ml • Purity of RNA sample = (A – A )/( A – A Thermo Fisher Scientific Thermo Scientific KingFisher Pure RNA Blood Kit...

  • Page 27: General Information

    RNA yield. The modifications of the elution step must be done in BindIt Software 3.2 and according to the volume ranges suitable for the KingFisher instrument. The table below indicates the available elution volumes of the KingFisher instruments. Thermo Fisher Scientific Thermo Scientific KingFisher Pure RNA Blood Kit...

  • Page 28: Decontamination And Disinfection Of Sample Material

    Use a decontaminant, such as Virkon™, paying due attention to the manufacturer’s instructions. You should also take care of the appropriate treatment and/or disposal of waste. Thermo Scientific KingFisher Pure RNA Blood Kit Thermo Fisher Scientific...

  • Page 29: Appendix A Troubleshooting

    Prolonged storage of the sample material may reduce the total RNA yield. Use only Thermo Scientific plates, strips, and tip combs with the KingFisher instruments. Use of products from other manufacturers may cause unsuitable mixing and affect the yield of purified RNA.
  • Page 30 RNA has already been released from the KingFisher Magnetic Beads into the Elution Buffer. If the KingFisher magnetic particle processor does not work properly, refer to the relevant user manual of the KingFisher instrument in use. Thermo Scientific KingFisher Pure RNA Blood Kit Thermo Fisher Scientific...

  • Page 31: Appendix B Typical Rna Yields

    Stabilized (PAXgene, Tempus) human blood 9−12 Human bone marrow 15−80 Fresh mouse blood 120−160 Fresh rabbit blood 25−30 Fresh canine blood 11−15 Fresh animal (horse, cow, sheep, goat) blood 2−5 Thermo Fisher Scientific Thermo Scientific KingFisher Pure RNA Blood Kit...

  • Page 33: Appendix C Ordering Information

    50 pcs 95040460 Microtiter deep well 96 plate, sterile 50 pcs 95040470 KingFisher Flex 24 deep well plate 50 pcs 95040480 KingFisher Flex 24 deep well plate, sterile 50 pcs Thermo Fisher Scientific Thermo Scientific KingFisher Pure RNA Blood Kit...
  • Page 34 KingFisher Flex 24 deep well plate, sterile 50 pcs 97003530 KingFisher Duo Combi pack for Microtiter deep well 1 box 96 plate (tips combs, plates, and elution strips for 96 samples) Thermo Scientific KingFisher Pure RNA Blood Kit Thermo Fisher Scientific...

  • Page 35: Thermo Fisher Scientific Thermo Scientific Kingfisher Pure Rna Blood Kit

    Chapter C General Information Thermo Fisher Scientific Thermo Scientific KingFisher Pure RNA Blood Kit...
  • Page 36 www.thermoscientific.com © 2013 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries. Specifications, terms and pricing are subject to change. Not all products are available in all countries. Please consult your local sales representative for details. Thermo Fisher Scientific 81 Wyman Street Waltham, MA 02451...

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