Instructions For Total Rna Purification From Paxgene Stabilized Blood Samples - Thermo Scientific KingFisher Pure RNA Blood Kit Instruction Manual

Thermo Fisher Scientific
When the protocol is completed, remove the plate and elution strip according
to the instructions on the KingFisher Duo display and switch off the
instrument. The purified RNA is ready for use in downstream applications.
When working with RNA, keep the purified samples on ice. Store the purified
RNA at -20°C or -80°C.
NOTE: The final RNA concentration in the nuclease-free water may
increase if the purified RNA is eluted into a smaller than recommended
volume of water, but this can slightly reduce the overall RNA yield.
Instructions for Total RNA Purification
from PAXgene Stabilized Blood Samples
1. Collect and store the blood samples in PAXgene tubes according to the
manufacturer's instructions.
2. If the PAXgene samples were stored in the cold, equilibrate the samples
at room temperature before purification according to the manufacturer's
instructions for approx. 2 h.
3. Centrifuge the PAXgene blood RNA tubes for 10 min at 3,000−5,000 x g.
4. Carefully discard the supernatant from each tube and wash the pellet.
Add 1−3 ml of nuclease-free water, vortex for 20−30 s, and collect the
pellet again by centrifuging for 10 min at 3,000–5,000 x g.
• For 0.5−2 ml of PAXgene blood lysate, wash the obtained pellet with
1 ml of water.
• For a whole tube of PAXgene blood lysate, wash the obtained pellet with
3 ml of water.
5. Carefully discard the supernatant, add 100 µl of nuclease-free water,
and vortex for 10−20 s to suspend precipitates. Add 400 µl of Lysis
Buffer (supplemented with DTT) and 20 µl of Proteinase K. Mix the lysate
and continue the purification as in
96 Deep Well Plates"
Duo with 12-pin Magnet Head"
Protocols and Pipetting Instructions
"Instructions for KingFisher Flex with
on page 18 or as in "Instructions for KingFisher
on page 20.
Thermo Scientific KingFisher Pure RNA Blood Kit
Chapter 5
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