Thermo Scientific KingFisher Plant DNA Kit Instruction Manual
Thermo Scientific KingFisher
Plant DNA Kit
Instruction Manual
Rev. 1.1
Thermo Scientific KingFisher
Plant DNA Kit
Instruction Manual
Rev. 1.1, Cat. no. N11996
Copyright © 2011 ermo Fisher Scientic Inc. First edition published in 2010. All rights reserved.
Reproduction of the accompanying user documentation in whole or in part is prohibited.
“BindIt”, “KingFisher”, “Microtiter”, “Multidrop”, “NanoDrop” and "Versette" are registered
trademarks of ermo Fisher Scientic.
All other trademarks and registered trademarks are the property of their respective holders.
ermo Fisher Scientic reserves the right to change its products and services at any time to
incorporate technological developments. is manual is subject to change without prior notice as part
of a continuous product development. Although this manual has been prepared with every precaution
to ensure accuracy, ermo Fisher Scientic assumes no liability for any errors or omissions, nor
for any damages resulting from the application or use of this information. is instruction manual
supersedes all previous editions.
e Product will operate substantially in conformance with ermo Fisher Scientic’s published
specications.
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ermo Fisher Scientic and its aliates shall have no liability to an End User arising out of the
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Thermo Fisher Scientific 5Thermo Scientific KingFisher Plant DNA Kit Instruction Manual
Contents
Chapter 1 Kit Content ...........................................................................................7
Chapter 2 Product Description.........................................................................11
Introduction ............................................................................11
Intended use .........................................................................11
Principle and procedure ........................................................11
Kit specications ......................................................................12
KingFisher magnetic particle processors ...................................13
Chapter 3 Safety Information............................................................................17
Chapter 4 Storage Conditions and Preparation of
the Working Solution ......................................................................19
Storage conditions ...................................................................19
Preparation of the RNaseA working solution ..........................19
Chapter 5 Protocols and Pipetting Instructions ...........................................21
Handling of KingFisher Magnetic Beads ..................................21
Sample material and storage .....................................................21
Homogenization of sample material .........................................22
Instructions for KingFisher Flex with 96 deep well plates
for DNA purication from 400µl of plant lysate .....................23
Summary of platecontents ...................................................25
Instructions for KingFisher Duo with 12-pin magnet head
and 96 deep well plates for DNA purication from
400µl of plant lysate ...............................................................26
Summary of plate and elution strip contents .........................28
Quantication and determination of the purity of DNA .........29
Chapter 6 General Information.........................................................................31
Reagent specicity and volumes ...............................................31
Handling of magnetic beads ....................................................31
Binding and wash steps ............................................................31
Elution step .............................................................................32
Thermo Fisher Scientific
Contents
6 Thermo Scientific KingFisher Plant DNA Kit Instruction Manual
Appendix A Troubleshooting ................................................................................35
Appendix B Ordering Information .......................................................................37
Thermo Fisher Scientific 7Thermo Scientific KingFisher Plant DNA Kit Instruction Manual
Chapter 1
Kit Content
Table 1-1. Thermo Scientific KingFisher Plant DNA Kit
Item KingFisher Plant DNA Kit
Cat. No. 97050196
Package size 1 x 96 samples
KingFisher Magnetic Beads 3.2 ml
Lysis Buffer 55 ml
Binding Buffer 50 ml
Wash Buffer 1 65 ml
Wash Buffer 2 65 ml
Wash Buffer 3 65 ml
Elution Buffer 20 ml
RNase A (lyophilized) 1 vial
Recommended instrument KingFisher Flex with
96 deep well head
KingFisher Duo
KingFisher 96
e KingFisher Plant DNA Kit (Cat. No. 97050196)
is intended for the purication of 96 samples using the
ermo Scientic KingFisher Flex with a 96 deep well
head or the ermo Scientic KingFisher Duo and
a lysed sample volume of 400µl.
Thermo Fisher Scientific8 Thermo Scientific KingFisher Plant DNA Kit Instruction Manual
Kit Content
Equipment and reagents to be supplied by the user:
• 80% ethanol (EtOH), molecular biology grade
• DNase-free water
• Homogenization equipment
• Magnetic particle processor
Table 1-2. Thermo Scientific KingFisher magnetic particle processors
Cat. No. Product
5400000 KingFisher magnetic particle processor
5400050 KingFisher mL magnetic particle processor
5400100 KingFisher Duo magnetic particle processor
5400630 KingFisher Flex magnetic particle processor with 96 deep well head
5400640 KingFisher Flex magnetic particle processor with 24 deep well head
Discontinued KingFisher 96 magnetic particle processor
Table 1-3. Thermo Scientific KingFisher Flex consumables
Cat. No. Product Package size
97002514 KingFisher Flex 96 tip comb for PCR magnet 80 pcs
97002524 KingFisher Flex 96 tip comb for KF magnet 100 pcs
97002534 KingFisher Flex 96 tip comb for deep well magnet 100 pcs
97002610 KingFisher Flex 24 deep well tip comb and plate 50 pcs
97002540 KingFisher Flex 96 KF plate (200 µl) 48 pcs
95040450 Microtiter deep well 96 plate, non sterile 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 9Thermo Scientific KingFisher Plant DNA Kit Instruction Manual
Kit Content
Table 1-4. Thermo Scientific KingFisher Duo consumables
Cat. No. Product Package size
97003500 KingFisher Duo 12-tip comb for Microtiter deep well 96 plate 50 pcs
97003510 KingFisher Duo 6-tip combs and KingFisher 24 deep well plate
(12 pcs of 24 deep well plates, each including 4 tips combs)
48 pcs
97003520 KingFisher Duo elution strip 40 pcs
97003530 KingFisher Duo Combi pack for Microtiter deep well 96 plate
(tips combs, plates and elution strips for 96 samples)
1 box
Table 1-5. Thermo Scientific KingFisher mL consumables
Cat. No. Product Package size
97002111 KingFisher mL tip comb 800 pcs
97002121 KingFisher mL tube 20 x 45 pcs
97002131 KingFisher mL combi (tubes and tip combs for 60 samples) 60
97002141 KingFisher mL combi (tubes and tip combs for 240 samples) 240
Table 1-6. Thermo Scientific KingFisher consumables
Cat. No. Product Package size
97002070 KingFisher tip comb 50 pcs
97002080 KingFisher plate 100 µl 50 pcs
97002084 KingFisher plate 200 µl 50 pcs
97002090 KingFisher plastics 100 µl 8-pack, 8 plates and 8 tip combs 1 box
97002094 KingFisher plastics 200 µl 8-pack, 8 plates and 8 tip combs 1 box
Thermo Fisher Scientific10 Thermo Scientific KingFisher Plant DNA Kit Instruction Manual
Kit Content
Thermo Fisher Scientific 11Thermo Scientific KingFisher Plant DNA Kit Instruction Manual
Chapter 2
Product Description
e KingFisher Plant DNA Kit is designed for rapid
automated purication of DNA from plant materials
using KingFisher instruments. e DNA puried with the
KingFisher Plant DNA Kit is of high quality and free of
proteins, nucleases, and other contaminants or inhibitors.
It is therefore suitable for direct use in many dierent
downstream applications, such as PCR (polymerase
chain reaction), restriction analysis and in several other
enzymatic reactions.
e KingFisher Plant DNA Kit is developed for
the purication of DNA from plant material using
paramagnetic particles. e purication process requires
no phenol/chloroform extraction or alcohol precipitation
and needs very little hands-on time. e reagents and
specic plastic consumables are designed to work with
the KingFisher Flex or KingFisher Duo magnetic particle
processors as part of an integrated system. e KingFisher
Plant DNA Kit is only intended for research use, not
for clinical or diagnostic use. e user is responsible for
validating the performance of the KingFisher instrument
and the KingFisher Plant DNA Kit for any particular use,
because the performance of the kits has not been validated
for any specic organism.
e KingFisher Plant DNA Kit uses magnetic-particle
technology for DNA purication. e ermo Scientic
KingFisher technology combines the speed and eciency
of DNA purication with easy handling of magnetic
particles. It is recommended that fresh, frozen, dried
Introduction
Intended use
Principle and
procedure
Thermo Fisher Scientific12 Thermo Scientific KingFisher Plant DNA Kit Instruction Manual
Product Description
Kit specifications
or lyophilized sample is mechanically disrupted into a
ne powder before the purication can start to ensure a
good yield of puried DNA. e powder is resuspended
into the Lysis Buer, mixed and incubated and then
sedimented after a short centrifugation step. e cleared
lysates are transferred to ermo Scientic KingFisher
plates for processing with a ermo Scientic KingFisher
magnetic particle processor. DNA binds to the surface
of the ermo Scientic KingFisher Magnetic Beads in
the presence of a chaotropic salt. e following eective
wash steps dispose of proteins, cell debris and any residual
contaminants, while the DNA bound to the KingFisher
Magnetic Beads is transferred through the wash steps.
First, two dierent Wash Buers are used in sequential
wash steps, followed by an ethanol wash. A rapid rinse in
the third Wash Buer removes the residual ethanol and
considerably improves the purity of the DNA. Highquality DNA is eluted into the Elution Buer and is ready
for subsequent downstream processes, such as enzymatic
reactions. Heat incubation is included in the elution step
when using the KingFisher Flex or KingFisher Duo.
e KingFisher Plant DNA Kit is designed for rapid
automated preparation of highly pure DNA from plant
materials using KingFisher magnetic particle processors.
e approximate processing time is less than 30 minutes
for the purication of 96 samples in the KingFisher Flex
and the KingFisher Duo. e obtained DNA can be
directly used in various downstream applications.
Plant samples can be fresh, frozen, lyophilized or dried.
Asuitable amount of fresh material is 20–50 mg. For
dried material, the amount of sample should be one fth
of the amount of fresh sample. e yields of acquired
puried DNA depend on the sample type, the method of
sample collection, and the method of tissue disruption.
e KingFisher Plant DNA Kit can be processed at
room temperature excluding the homogenization with
Kit specifications
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