Thermo Scientific KingFisher Pure RNA Blood Kit, Инструкция по эксплуатации

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18 | Thermo Scientific KingFisher Pure RNA Blood Kit Thermo Fisher Scientific
Chapter 5 | Protocols and Pipetting Instructions
Instructions for KingFisher Flex with
96 Deep Well Plates
These instructions are intended for RNA purification from 200 µl of whole
blood, using the 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.
For each run, prepare the DNase I working solution and Lysis Buffer with
2 M DTT. For more instructions, refer to Chapter 4:
“Storage Conditions and
Preparation of the Reagents” .
Check all the solutions in the kit for salt precipitation before each use.
Redissolve precipitates by warming the solution at 37°C and equilibrate to
room temperature (15−25°C).
Blood samples of certain species, e.g. mouse, contain a high amount of
RNA. With such species, it is recommended to use up to 50 µl of blood
sample to obtain RNA of equal yield and purity. When working with less than
a 200 µl volume of blood sample, it is recommended to equalize the volume
by adding Lysis Buffer, thus bringing the total volume to 400 µl.
1. Take six empty Microtiter deep well 96 plates and two empty Thermo
Scientific™ KingFisher™ Flex 96 KF plates.
2. Fill the
plates
as follows.
Plate
number
Plate type Plate name Content
Reagent volume
per well
1
Microtiter deep
well 96 plate
Sample Lysis Buffer
Blood sample
Proteinase K
200 µl
200 µl
20 µl
2 Wash 1_1 Wash Buffer 1 900 µl
3 DNase DNase I working
solution 70 µl
4 Wash 1_2 Wash Buffer 1 700 µl
5 Wash 2_1 Wash Buffer 2 700 µl
6 Wash 2_2 Wash Buffer 2 700 µl
7 KingFisher Flex
96 KF plate Elution Nuclease-free
water 75 µl