Thermo Scientific™ GeneJET Endo-Free Plasmid Maxiprep Kit

Catalog No.	Quantity
FERK0861	10 Preps

Catalog No. FERK0861 Supplier Thermo Scientific™ Supplier No. K0861

Description

Includes

Resuspension Solution, Lysis Solution, Neutralization Solution, Endotoxin Binding Reagent, Wash Solution I (concentrated), Wash Solution II (concentrated), RNase A Solution, Elution Buffer (10mM Tris-HCl, pH 8.5), GeneJET Maxi Purification Columns pre-assembled with collection tubes (50mL) and Collection tubes (50mL).

The Thermo Scientific GeneJET Plasmid Maxiprep Kit utilizes silica-based membrane technology in the form of convenient spin column.

Thermo Scientific GeneJET Endo-Free Plasmid Maxiprep Kit is a fast, convenient, and efficient system for the large scale purification of endotoxin-free plasmid DNA. Endotoxins are major contaminants found in plasmid DNA preparations. For transfection of sensitive cell lines endotoxins are a major problem. The same applies to in vivo transfection as endotoxins can elicit strong immune responses. Thermo Scientific GeneJET Endo-Free Plasmid Maxiprep Kit is designed to reduce endotoxin level to <0.1 EU/μg of plasmid DNA. The kit utilizes a silica-based membrane technology in the form of convenient spin columns.

Each preparation recovers up to 1000 μg of high copy plasmid DNA from 350 mL of recombinant E. coli cultures that can be used in a wide variety of molecular biology procedures such as transfection of sensitive cells (e.g. Huh-7), in vivo transfection, transformation, restriction endonuclease digestion, automated sequencing, and in vitro transcription.

Highlights

• Pure—free of endotoxins plasmid DNA suitable for transfection of sensitive cells
• Efficient—plasmid recovery resulting in high DNA yields
• Fast—isolation of high quality plasmid DNA in less than 60 minutes
• Convenient—spin column format, centrifuge or vacuum protocol

Applications

Isolated plasmid DNA is suitable for all conventional molecular biology procedures, including:

• Transfection of sensitive cell lines
• FastDigest restriction enzyme digestion
• In vivo transfection
• Automated fluorescent sequencing
• PCR
• Cloning
• Transformation
• In vitro transcription

Includes

• Resuspension Solution
• Lysis Solution
• Neutralization Solution
• Endotoxin Binding Reagent
• Wash Solution I (concentrated)
• Wash Solution II (concentrated)
• RNase A Solution
• Elution Buffer (10 mM Tris-HCl, pH 8.5)
• GeneJET Endo-Free Maxi Purification Columns & Collection Tubes (50 mL, blue-capped)
• GeneJET Endo-Free Maxi Filtration Columns & Collection Tubes (50 mL, white-capped)

Hazardous: No

Quality Control:

• The GeneJET Plasmid Maxiprep Kit is qualified by isolating high copy number plasmid DNA from 250 mL of overnight E. coli culture (OD600 = 2) grown in LB medium following the protocol outlined in the manual.
• The kit passes QC requirements if the purified plasmid DNA has an A_260/280 ratio between 1.8 and 1.9 and a dominant band of supercoiled plasmid DNA is observed after agarose gel electrophoresis.
• The functional quality of the plasmid DNA is evaluated by digestion with restriction endonucleases.

Order Info

Orders placed Monday through Wednesday ship next day; Thursday and Friday orders ship the following Monday.

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Specifications

• Concentration: 400 ng/μL
• Column Type: Spin Column
• Format: Spin Column
• Isolation Technology: Silica Column
• Scale: Maxi
• Sample Type: Bacteria
• For Use With (Application): Real-Time Quantitative PCR (qPCR), Transfection, Cloning, Sequencing, In Vitro Transcription, Transformation, Nucleic Acid Labeling, PCR
• High-throughput Compatibility: Not High-throughput Compatible (Manual)
• No. of Reactions: 10 Preps
• Prep Scale: > 200 μg (Large-Scale) Plasmid DNA
• Product Line: GeneJET
• Product Type: Endo-Free Plasmid MaxiPrep Kit
• Quantity: 10 Preps
• Target: Plasmid DNA
• Test Time: 60 min.

Frequently Asked Questions (FAQs)

Can I use water instead of Elution Buffer in the elution step of GeneJET and MagJET nucleic acid purification kits?

We would highly recommend using the provided Elution Buffer in the elution step for best results. You may use nuclease-free water; however, it may affect the yield of the nucleic acids during purification and their stability during storage.

Are MagJET and GeneJET plasmid purification kits compatible with EndA+ bacterial strains?

Yes, MagJET and GeneJET Plasmid DNA Purification Kits are suitable for plasmid DNA purification from EndA+ strains. Please refer to the specific kit protocol for details. GeneJET Plasmid Miniprep Kit requires Wash Solution I (Cat. No. R1611, not included in the kit) for EndA+ strains.

Can I use a vacuum manifold with GeneJET kits?

The GeneJET spin columns are compatible with vacuum manifolds of standard size (e.g. Qiagen, Sigma-Aldrich, Promega). However, not all GeneJET kits are supported with a vacuum protocol. Please refer to the particular kit protocol for the detailed procedure.

At what temperature should I perform centrifugation steps in the GeneJET kits?

We recommend performing centrifugation steps at room temperature when the temperature is not specified.

Are GeneJET spin columns or MagJET magnetic beads available separately?

No, we do not offer GeneJET columns or MagJET magnetic beads separately at this time.

How can I increase plasmid DNA yield during elution from GeneJET plasmid kits?

To increase the overall plasmid DNA yield by 20-30%, additional elution steps may be performed.

Can I scale-up culture volume for the GeneJET Endo-free Plasmid Maxiprep Kit?

Generally, 350 mL of overnight bacterial culture grown in LB medium is sufficient for a good yield of high-copy and low-copy plasmid samples. However, it is important not to exceed the recommended cell mass (culture volume X OD600) because it may decrease the quality of isolated DNA. The maximum culture volume to use can be determined using the formula: Maximum culture volume (mL) = 750/OD600.

Which culture media are recommended for bacteria to be used with GeneJET Plasmid Kits? Can I use a rich growth medium?

The GeneJET Plasmid purification protocols are optimized for bacterial cultures grown in Luria-Bertani (LB) medium. The use of rich growth media is not recommended.

Which bacterial strains can be used for plasmid purification with GeneJET Plasmid KitS?

High quality plasmid DNA can be obtained from various E. coli strains including DH10B, DH5a, XL1-Blue, JM109, JM107, and TOP10.

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov

For Research Use Only. Not for use in diagnostic procedures.