Thermo Fisher Scientific ChargeSwitch Plasmid ER Mini Kit User guide

Brand
Thermo Fisher Scientific
Model
ChargeSwitch Plasmid ER Mini Kit
Type
User guide
For Research Use Only. Not for use in diagnostic procedures.
ChargeSwitch™ Plasmid ER Mini Kit
USER GUIDE
For purification of plasmid DNA (endotoxin-reduced) from
bacterial cells
Catalog Number CS10100
Publication Number MAN0025683
Revision A.0
Life Technologies Corporation | 5781 Van Allen Way | Carlsbad, California 92008 USA
For descriptions of symbols on product labels or product documents, go to thermofisher.com/symbols-definition.
The information in this guide is subject to change without notice.
DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S) WILL NOT BE
LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR
ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.
Revision history: Pub. No. MAN0025683
Revision Date Description
A.0 15 October 2021 • Updated publication format: MAN0025683 Rev. A.0 supersedes
previous publication - Cat. No. CS10100, Version B, 2003.
• Added new procedure for the kit - Chapter 4, “Automated method”.
Important Licensing Information: These products may be covered by one or more Limited Use Label Licenses. By use of these
products, you accept the terms and conditions of all applicable Limited Use Label Licenses.
NOTICE TO PURCHASER: DISCLAIMER OF LICENSE: Purchase of this software product alone does not imply any license under any
process, instrument or other apparatus, system, composition, reagent or kit rights under patent claims owned or otherwise controlled by
Thermo Fisher Scientific, either expressly, or by estoppel.
TRADEMARKS: All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified.
©2021 Thermo Fisher Scientific Inc. All rights reserved.
Contents
■CHAPTER 1 Product information .................................................. 5
Product description ............................................................. 5
Advantages of the ChargeSwitch™ Plasmid ER Mini Kit .......................... 5
Overview of the ChargeSwitch™ technology .................................... 5
Kit specifications ........................................................... 6
Contents and storage ............................................................ 6
Required materials not supplied ................................................... 7
■CHAPTER 2 Before you begin ..................................................... 8
Procedural guidelines ............................................................ 8
Guidelines for elution buer .................................................. 8
Guidelines for magnetic bead handling ........................................ 8
Guidelines for bacterial cultures .............................................. 9
Before you begin ................................................................ 9
■CHAPTER 3 Manual method ..................................................... 10
Lyse bacteria .................................................................. 10
Bind DNA ..................................................................... 10
Wash DNA .................................................................... 11
Elute DNA ..................................................................... 11
■CHAPTER 4 Automated method ................................................. 13
Lyse bacteria .................................................................. 13
Prepare the processing plates ................................................... 14
Process samples on the instrument .............................................. 14
■APPENDIX A Determine plasmid DNA yield and quality ....................... 15
Calculate plasmid DNA yield with UV absorbance .................................. 15
Quant-iT™ kits ................................................................. 15
Plasmid DNA quality ............................................................ 15
ChargeSwitch™ Plasmid ER Mini Kit User Guide 3
■APPENDIX B Accessory products ............................................... 16
Accessory products ............................................................ 16
E‑Gel™ Agarose Gels and DNA ladders ........................................... 16
MagnaRack™ Magnetic Separation Rack .......................................... 17
■APPENDIX C Troubleshooting .................................................... 18
■APPENDIX D Safety ............................................................... 20
Chemical safety ................................................................ 21
Biological hazard safety ......................................................... 22
■APPENDIX E Documentation and support ...................................... 23
Customer and technical support ................................................. 23
Limited product warranty ........................................................ 23
Contents
4ChargeSwitch™ Plasmid ER Mini Kit User Guide
Product information
IMPORTANT! Before using this product, read and understand the information in the “Safety” appendix
in this document.
Product description
The ChargeSwitch™ Plasmid ER Mini Kit allows rapid and ecient purification of highly pure
plasmid DNA from E. coli cells. This protocol guides users through cell lysate preparation using the
modified alkaline lysis procedure, followed by manual or automated plasmid DNA purification using
ChargeSwitch™ technology. Automated plasma DNA purification is performed using the KingFisher™
Flex instrument.
The kit includes Endotoxin Reduction Reagent (ETRR) that lowers the endotoxin levels (<50 EU / μg
DNA) in the purified plasmid DNA.
Advantages of the ChargeSwitch™ Plasmid ER Mini Kit
•High yield—Isolates up to 20 μg of plasmid DNA from E. coli using a magnetic bead‑based
technology without the use of chaotropic salts, organic solvents, or ethanol.
•Fast protocol—Isolates plasmid DNA from six samples in ~ 20 minutes using multi-channel
pipettors and MagnaRack™ (manual procedure) or up to 96 samples in 22 minutes using the
KingFisher™ Flex Magnetic Bead Processor (KingFisher™ procedure).
•High quality and reliable performance—Provides purified plasmid DNA in a variety of
applications including mammalian cell transfection, automated and manual sequencing,
amplification reactions, bacterial cell transformation, cloning, and labeling.
Overview of the ChargeSwitch™ technology
The ChargeSwitch™ technology is a novel magnetic bead‑based technology that provides a switchable
surface which is charge-dependent on the pH of the surrounding buer to help with nucleic acid
purification.
In low pH conditions, the ChargeSwitch™ Magnetic Beads have a positive magnetic charge that binds
the negatively charged nucleic acid backbone. Proteins and other contaminants are not bound and are
washed away in aqueous buers. To elute the nucleic acids, the charge on the surface is neutralized by
raising the pH to 8.5 using a low salt elution buer. Purified DNA elutes instantly and is ready for use in
downstream applications.
1
ChargeSwitch™ Plasmid ER Mini Kit User Guide 5
Kit specifications
Table 1 Specifications of the ChargeSwitch™ Plasmid ER Mini Kit
Specifications ChargeSwitch™ Plasmid ER Mini Kit
Starting material 1–5 mL fresh, overnight LB culture
Bead binding capacity 1 mg beads bind ~25 μg plasmid DNA
Bead size 1 μm
Bead concentration 25 mg/ml
Bead storage buer 10 mM MES, pH 5.0
10 mM NaCl, 0.1% Tween™ 20
Elution volume 50–100 μL
DNA Yield Up to 20 μg
Contents and storage
All components of the ChargeSwitch™ Plasmid ER Mini Kit are shipped at room temperature. Upon
receipt, store as indicated in Table 2.
Reagents provided in the kit are sucient for 50 preps using 1 mL bacterial culture input.
IMPORTANT! Do not freeze the magnetic beads as freezing damages the beads. Beads that have
been frozen cannot be used for nucleic acid purifications.
Table 2 ChargeSwitch™ Plasmid ER Mini Kit (Cat. No. CS10100)
Content Amount[1] Storage[2] Stability
ChargeSwitch™ Precipitation Buer (N5) 15 mL 4°C 12 months
RNase A (5 mg/ml in 10 mM Tris-HCl, pH 8.5,
10 mM EDTA)
0.4 mL
Resuspension Buer (R4) (R4; 10 mM Tris-
HCl, pH 8.5, 10 mM EDTA)
15 mL
ChargeSwitch™ Magnetic Beads (25 mg/ml in
10 mM MES, pH 5.0, 10 mM NaCl, 0.1%
Tween™ 20)
2 x 1 mL 18 to 25°C
ChargeSwitch™ Lysis Buer (L9) 15 mL
ChargeSwitch™ Wash Buer (W11) 50 mL
ChargeSwitch™ Wash Buer (W12) 50 mL
Chapter 1 Product information
Contents and storage
1
6ChargeSwitch™ Plasmid ER Mini Kit User Guide
Table 2 ChargeSwitch Plasmid ER Mini Kit (Cat. No. CS10100) (continued)
Content Amount[1] Storage[2] Stability
ChargeSwitch™ Elution Buer (E5) 10 mL 18 to 25°C 12 months
Endotoxin Reduction Reagent (D1 / ETRR) 4.5 mL
[1] Some components may have higher volumes than listed in the table.
[2] See product label for expiration date.
Required materials not supplied
Unless otherwise indicated, all materials are available through thermofisher.com. "MLS" indicates that
the material is available from fisherscientific.com or another major laboratory supplier.
Catalog numbers that appear as links open the web pages for those products.
Item Source
Automated nucleic acid extraction system and materials
KingFisher™ Flex Magnetic Particle Processor with 96 deep‑well
magnet 5400630
V or round bottom 96 DeepWell™ plate for pellet preparation MLS
KingFisher™ Flex Microtiter Deep-Well 96 plate ("KF DeepWell™") 95040450 or 95040460
96 Standard-Well Plates for KingFisher™ Flex Magnetic Particle
Processor ("KF Standard") 97002540
96 Deep-Well Tip Combs for KingFisher™ Flex Magnetic Particle
Processor 97002534
Manual nucleic acid extraction system and materials
MagnaRack™ Magnetic Separation Rack CS15000
Sterile 1.5 mL microcentrifuge tubes MLS
Equipment
Adjustable micropipettors MLS
Adjustable‑speed rotating platform shaker (optional) MLS
Chapter 1 Product information
Required materials not supplied 1
ChargeSwitch™ Plasmid ER Mini Kit User Guide 7
Before you begin
Procedural guidelines
Follow the recommendations below to obtain the best results.
• Perform all steps at room temperature (18–25°C) unless otherwise noted.
• Maintain a sterile environment when handling DNA to avoid any contamination from DNases.
• Ensure that no DNase is introduced into the solutions supplied with the kit.
• Ensure all equipment coming in contact with DNA is sterile, including pipette tips and tubes.
• Perform the centrifugations during cell lysis at 4°C to improve the eciency of the precipitation
reaction.
• Perform the recommended wash steps during the purification to obtain the best results.
Guidelines for elution buer
IMPORTANT! Do not use water for elution.
The plasmid DNA is eluted with Elution Buer (E5; 10 mM Tris‑HCl, pH 8.5). To obtain the best results,
always use Elution buer (E5) to elute the DNA. If another buer is used to elute the plasmid DNA, it
must have a pH of 8.5–9.0. Buers with a pH <8.5 will not elute plasmid DNA.
The plasmid DNA is eluted with 50–100 μL of elution buer. The volume of elution buer can be
changed to obtain plasmid DNA in the desired final concentration. To obtain best results, always use
elution buer that is equal or greater than the volume of beads used in the protocol. If the volume
of elution buer is lower than the volume of beads, DNA elution is incomplete, and you may need to
perform a second elution to recover all DNA.
Guidelines for magnetic bead handling
Follow the guidelines for the best results.
IMPORTANT! Do not freeze the magnetic beads as freezing damages the beads. Beads that have
been frozen cannot be used for nucleic acid purifications.
• Pipette up and down gently to mix beads and avoid forming bubbles.
• Do not allow beads to dry; this reduces bead binding ecacy.
• To aspirate the supernatant after bead washing, place the pipette tip away from the pellet to
carefully remove the supernatant without disturbing or removing any beads.
2
8ChargeSwitch™ Plasmid ER Mini Kit User Guide
Guidelines for bacterial cultures
Follow the guidelines for growing bacterial cultures to obtain the best results.
•To obtain the best results, use fresh, overnight cultures. The kit can also be used to purify plasmid
DNA from frozen cell pellets.
•Grow transformed E. coli in LB medium with the appropriate antibiotic. If desired, a richer medium
can be used to grow E. coli.
•Use 1–5 mL of overnight bacterial cultures with an absorbance at 600 nm (A600) of 9 OD.
Before you begin
• Prepare the Resuspension Buer (R4): Add all of the supplied RNase A to the Resuspension Buer
(R4), then mix. Mark the bottle label to indicate addition of RNase A. Store prepared Resuspension
Buer (R4) at 4°C.
• Chill an aliquot of Precipitation Buer (N5) to 4°C.
• Check the Lysis Buer (L9) for precipitates. If precipitates are present, warm the solution briefly at
37°C to dissolve the precipitate.
Chapter 2 Before you begin
Before you begin 2
ChargeSwitch™ Plasmid ER Mini Kit User Guide 9
Manual method
Lyse bacteria
1. Centrifuge 1–5 mL of overnight bacterial culture at room temperature, then discard supernatant.
2. Add 150 μL of prepared Resuspension Buer (R4) containing RNase A to the cell pellet. Pipet up
and down to resuspend.
3. Add 150 μL of Lysis Buer (L9). Mix by inverting 6 times.
4. Incubate for 2–5 minutes at room temperature.
IMPORTANT! Do not incubate for longer than 5 minutes.
5. Add 150 μL of chilled Precipitation Buer (N5). Mix by inversion until a white, free‑flowing,
precipitate is formed.
6. Centrifuge tube in a microcentrifuge at maximum speed for 10 minutes at 4°C.
7. Transfer clear supernatant to a new sterile microcentrifuge tube.
Bind DNA
1. Vortex ChargeSwitch™ Magnetic Beads to fully resuspend and evenly distribute the beads in the
storage buer.
2. (Optional) Add 90 μL of Endotoxin Reduction Reagent (D1) to the samples if low endotoxin level is
desirable.
3. Add 40 μL of ChargeSwitch™ Magnetic Beads to the lysate.
4. Mix by pipetting gently up and down without forming bubbles.
5. Incubate at room temperature for 1 minute to allow the DNA to bind to the beads.
6. Place the sample on the MagnaRack™ Magnetic Separation Rack for 1 minute or until the beads
have formed a tight pellet.
7. Without removing the tube from the magnetic rack, remove and discard supernatant. Avoid
disturbing the beads by angling the pipette tip away from the pellet.
8. Proceed immediately to wash DNA (next section).
3
10 ChargeSwitch™ Plasmid ER Mini Kit User Guide
Wash DNA
1. Remove the tube containing the pelleted magnetic beads from the MagnaRack™ Magnetic
Separation Rack.
2. Add 1 mL of Wash Buer (W11) to the tube. Pipet up and down gently to mix the sample without
forming bubbles.
3. Place the sample on the MagnaRack™ Magnetic Separation Rack for 1 minute or until the beads
have formed a tight pellet.
4. Without removing the tube from the magnetic rack, remove and discard supernatant. Avoid
disturbing the beads by angling the pipette tip away from the pellet.
5. Add 1 mL of Wash Buer (W12) to the tube. Pipet up and down gently to mix the sample without
forming bubbles.
6. Place the sample on the MagnaRack™ Magnetic Separation Rack for 1 minute or until the beads
have formed a tight pellet.
7. Without removing the tube from the magnetic rack, remove and discard supernatant. Avoid
disturbing the beads by angling the pipette tip away from the pellet.
8. Proceed immediately to elute DNA (next section).
Elute DNA
1. Remove the tube containing the pelleted magnetic beads from the MagnaRack™ Magnetic
Separation Rack.
2. Add 50–100 μL of Elution Buer (E5; 10 mM Tris‑HCl, pH 8.5) to the tube. Pipet up and down
gently to mix the sample without forming bubbles.
3. Incubate at room temperature for 1 minute.
(Optional) For maximum yield, halfway through incubation, mix the suspension of beads by
pipetting up and down gently.
4. Place the sample on the MagnaRack™ Magnetic Separation Rack for 1 minute or until the beads
have formed a tight pellet.
5. Without removing the tube from the magnetic rack, carefully transfer the supernatant containing
the purified plasmid DNA into a new, nuclease-free tube. Avoid aspirating the beads by angling
the pipette tip away from the pellet.
Note: If the supernatant containing the DNA is discolored, repeat step 4 and step 5.
6. Discard the used magnetic beads.
Note: Do not re‑use the magnetic beads.
Chapter 3 Manual method
Wash DNA 3
ChargeSwitch™ Plasmid ER Mini Kit User Guide 11
The purified plasmid DNA is ready for immediate use.
Note: Alternatively, the plate can be sealed and stored at -20°C.
Chapter 3 Manual method
Elute DNA
3
12 ChargeSwitch™ Plasmid ER Mini Kit User Guide
Automated method
Before you begin, download the KingFisher™ Flex CS10100_CS_Plasmid_ER_Mini_Kit.bdz script from
the CS10100 product page at www.thermofisher.com and load onto the KingFisher™ Flex instrument.
Lyse bacteria
Follow this lysis procedure for plasmid DNA purification with the KingFisher™ Flex Magnetic Particle
Processor with 96 deep‑well magnet.
1. Centrifuge 1–5 mL of overnight bacterial culture per well at room temperature, then discard
supernatant.
2. Add 150 μL of prepared Resuspension Buer (R4) containing RNase A to each cell pellet. Pipet up
and down to resuspend.
3. Add 150 μL of Lysis Buer (L9) per well. Mix by inverting 6 times.
4. Incubate for 2–5 minutes at room temperature.
IMPORTANT! Do not incubate for longer than 5 minutes.
5. Add 150 μL of chilled Precipitation Buer (N5) per well. Mix by inversion until a white, free‑flowing,
precipitate is formed in each well.
6. Centrifuge at maximum speed for 10 minutes at 4°C or until lysates have cleared.
7. Proceed immediately to prepare the processing plates (next section).
4
ChargeSwitch™ Plasmid ER Mini Kit User Guide 13
Prepare the processing plates
Prepare the plates according to Table 3.
Table 3 Processing plate configurations
Plate ID Plate Type Reagent Volume per well
Plasmid Plate KF DeepWell™ChargeSwitch™ Magnetic
Beads
30 μL
Endotoxin Reduction
Reagent (D1/ETRR)
45 μL
Wash Plate 1 KF DeepWell™Wash Buer (W11) 900 μL
Wash Plate 2 KF DeepWell™Wash Buer (W12) 900 μL
Elution Plate KF Standard Elution Buer (E5) 100 μL[1]
Tip comb KF Standard Place a tip comb in the plate.
[1] When purifying a low copy number plasmid, elution volume can be reduced (50 μL minimum).
Process samples on the instrument
1. Transfer 450 μL of supernatant prepared in step 6 of "Lyse bacteria" to the KF DeepWell™ plate
labeled "Plasmid Plate".
2. Select the CS10100_CS_Plasmid_ER_Mini_Kit.bdz script on the KingFisher™ Flex instrument and
start the run.
3. Follow the prompts on the instrument screen to load the plates and start the extraction.
4. At the end of the run, immediately remove the plate labeled "Elution Plate". Discard the other
plates.
The purified plasmid DNA is ready for immediate use. Alternatively, the plate can be sealed and
stored at -20°C.
Note: If insucient plasmid DNA is obtained, the lysate and ETRR volumes in the Plasmid Plate
can be adjusted as follows: Lysate 800 μL, ETRR 80 μL.
Chapter 4 Automated method
Prepare the processing plates
4
14 ChargeSwitch™ Plasmid ER Mini Kit User Guide
Determine plasmid DNA yield and
quality
Calculate plasmid DNA yield with UV absorbance
Perform DNA quantitation using UV absorbance at 260 nm.
Note: Use a cuvette with an optical path length of 1 cm blanked against 10 mM Tris‑HCl, pH 7.5.
1. Prepare a dilution of the DNA in 10 mM Tris‑HCl, pH 7.5. Mix well.
2. Measure the absorbance of the dilution at A260 in a spectrophotometer.
3. Calculate the concentration of DNA using the following formula:
DNA (μg/mL) = A260 ✕ 50 ✕ dilution factor
For DNA, A260 = 1 for a 50 μg/mL solution measured in a cuvette with an optical path length of
1 cm.
Quant-iT™ kits
The Quant-iT™ kits provide a rapid, sensitive, and specific fluorescent method for dsDNA quantitation.
The kit contains a state‑of‑the‑art quantification reagent, DNA standards for a standard curve, and
pre‑made buer to allow fluorescent DNA quantification using standard fluorescent microplate readers.
Plasmid DNA quality
Typically, plasmid DNA isolated using the ChargeSwitch™ Plasmid ER Mini Kit has an A260/ A280 ratio
1.7 or higher when samples are diluted in Tris‑HCl pH 7.5, indicating that the DNA is reasonably clean
of proteins that could interfere with downstream applications. Absence of contaminating RNA can be
confirmed by agarose gel electrophoresis.
A
ChargeSwitch™ Plasmid ER Mini Kit User Guide 15
Accessory products
Accessory products
Unless otherwise indicated, all materials are available through thermofisher.com. "MLS" indicates that
the material is available from fisherscientific.com or another major laboratory supplier.
Product Amount Source
MagnaRack™ Magnetic Separation Rack 1 rack CS15000
Quant-iT™ DNA Assay Kit, High Sensitivity 1000 assays Q33120
Quant-iT™ DNA Assay Kit, Broad-Range 1000 assays Q33130
Quant-iT™ PicoGreen™ dsDNA Reagent 1 kit, 1 mL P7589
Luria Broth Base (Miller's LB Broth Base), powder 2.5 kg 12795084
ChargeSwitch™ NoSpin Plasmid Micro Kit 96 reactions CS10201
ChargeSwitch™ gDNA Micro Tissue Kit 50 reactions CS11203
ChargeSwitch™ gDNA Mini Tissue Kit 25 reactions CS11204
ChargeSwitch™ gDNA Blood Kit, 50–100 µL 50 reactions CS11040
E‑Gel™ Agarose Gels and DNA ladders
E‑Gel™ Agarose Gels are buerless pre-cast agarose gels designed for fast, convenient electrophoresis
of DNA samples. The gels are available in dierent agarose percentage and well formats for your
convenience.
A large variety of DNA ladders are available from Thermo Fisher™ for sizing DNA.
For more details on these products, visit our website at www.thermofisher.com.
B
16 ChargeSwitch™ Plasmid ER Mini Kit User Guide
MagnaRack™ Magnetic Separation Rack
Use the MagnaRack™ Magnetic Separation Rack with the ChargeSwitch™ Plasmid ER Mini Kit to obtain
the best results. Other magnetic separators may not provide similar magnetic strength or may not be
compatible with the volumes used in the protocol.
The MagnaRack™ Magnetic Separation Rack, available from Thermo Fisher™ (Cat. No. CS15000), is a
two-piece magnetic separation rack for protocols with magnetic beads. It consists of a magnetic base
station and a removable tube rack. The tube rack fits onto the magnetic base station in two dierent
positions associating the row of 12 neodymium magnets with a single row of 12 tubes for simple and
easy ‘on the magnet’ and ‘o the magnet’ sample processing.
For more information, see www.thermofisher.com.
Appendix B Accessory products
MagnaRack™ Magnetic Separation Rack B
ChargeSwitch™ Plasmid ER Mini Kit User Guide 17
Troubleshooting
Observation Possible cause Recommended action
Low plasmid DNA yield Poor quality of starting material
or incomplete lysis
Check the growth conditions of the cell culture
to ensure plasmid propagation. Use a high
copy number plasmid if possible.
If the cell lysate is too viscous, reduce the
amount of cells used per sample.
Ensure complete resuspension of the bacterial
cell pellet. Decrease the amount of starting
material used.
Use Precipitation Buer (N5) chilled to 4°C and
perform centrifugation at 4°C to improve the
precipitation eciency and plasmid DNA yield.
Increase the incubation time during lysis but
do not exceed 5 minutes.
Non-optimal elution conditions After adding Elution Buer (E5) to the sample,
pipet up and down to resuspend the magnetic
beads before incubation.
Increase the mixing steps during elution to 20–
30 times.
If you are using a dierent buer for elution,
ensure the pH of the buer is 8.5–9.0.
Heat elution to 56°C for 8 minutes
before pipetting purified plasmid DNA
(manual procedure) or use KingFisher™
CS10100_Plasmid_heatedelution.bdz script
(available from Technical Support).
Magnetic beads not functional Do not freeze magnetic beads. Store the
magnetic beads at room temperature. Do not
re‑use the magnetic beads.
Incorrect magnetic bead
amount used for binding
Use the recommended amount of magnetic
beads for binding.
If plasmid DNA yields are lower, you may
increase the bead amount used.
Aspiration of beads during
pipette transfers
Ensure no beads are aspirated during removal
of buers (manual procedure).
Leftover beads in KingFisher™
plates
Ensure KingFisher™ instrument magnet is
properly aligned (automated procedure).
C
18 ChargeSwitch™ Plasmid ER Mini Kit User Guide
Observation Possible cause Recommended action
Eluate containing plasmid DNA
is discolored
Incomplete bead removal Place the sample in the MagnaRack™
Magnetic Separation Rack until the beads
form a tight pellet. Remove the eluate to a
sterile microcentrifuge tube, taking care not
to disturb the bead pellet (manual procedure).
Resource Technical Support for help with
optimizing KingFisher™ script parameters
(automated procedure).
Genomic DNA contamination Genomic DNA sheared during
handling
Gently invert tubes to mix after adding buers.
Do not vortex as it can shear genomic DNA.
To eciently precipitate the genomic DNA
away from the plasmid DNA, the genomic
DNA must be intact.
Plasmid DNA degradation Incorrect lysis procedure Incubate the lysate at room temperature for no
longer than 5 minutes.
Bubbles formed during mixing
steps
Make sure that the pipette tip is submerged in
the solution during mixing.
Appendix C Troubleshooting
MagnaRack™ Magnetic Separation Rack C
ChargeSwitch™ Plasmid ER Mini Kit User Guide 19
Safety
WARNING! GENERAL SAFETY. Using this product in a manner not specified in the user
documentation may result in personal injury or damage to the instrument or device. Ensure that
anyone using this product has received instructions in general safety practices for laboratories and
the safety information provided in this document.
·Before using an instrument or device, read and understand the safety information provided in the
user documentation provided by the manufacturer of the instrument or device.
·Before handling chemicals, read and understand all applicable Safety Data Sheets (SDSs) and use
appropriate personal protective equipment (gloves, gowns, eye protection, and so on). To obtain
SDSs, see the “Documentation and Support” section in this document.
D
20 ChargeSwitch™ Plasmid ER Mini Kit User Guide
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Thermo Fisher Scientific ChargeSwitch Plasmid ER Mini Kit User guide

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Thermo Fisher Scientific
Model
ChargeSwitch Plasmid ER Mini Kit
Type
User guide
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