Invitrogen™ PureLink™ Fast Low-Endotoxin Maxi Plasmid Purification Kit

Catalog No.	No. of Reactions
A36228	20 Preps

Catalog No. A36228 Supplier Invitrogen™ Supplier No. A36228

Description

The PureLink Fast Low-Endotoxin Maxi Plasmid Purification Kit uses next-generation columns with advanced silica membranes that enable isolation of transfection grade (<1 EU/μg) plasmid DNA in about 30 minutes.

The simple precipitation-free protocol yields up to 1.5 mg plasmid, suitable for standard transfections and all molecular biology applications such as cloning and sequencing.

• Ultra-fast protocol—innovative silica-based columns enable purification of transfection grade plasmid DNA in under 30 minutes
• Low endotoxin plasmid—the resulting plasmid contains less than 1 EU/μg endotoxins
• High yields—isolate up to 1.5 mg of high-quality plasmid DNA
• Colored buffers—the kit features colored buffers that permit error-free visualization of complete bacterial cell lysis and subsequent neutralization

Specifications

• Content And Storage: • Resuspension buffer, store at 4°C upon reciept
  • Lysis buffer, store at room temperature
  • Precipitation buffer, store at room temperature
  • Binding buffer, store at room temperature
  • Wash buffer 1, store at room temperature
  • Wash buffer 2 (concentrate), store at room temperature
  • Elution buffer, store at room temperature
  • Column assemblies, store at room temperature
  • Syringe filters, store at room temperature
  • Syringe plungers, store at room temperature
  • Collection tubes, store at room temperature
  
  
• Format: Kit
• Isolation Technology: Silica Column
• Scale: Maxi
• For Use With (Application): Cloning, PCR
• No. of Reactions: 20 Preps
• Prep Scale: Maxi input culture of 100-500 mL to obtain a yield of up to 850 μg
• Product Line: PureLink
• Product Type: Plasmid Purification Kit
• Quantity: 20 Preps
• Shipping Condition: Room Temperature
• Test Time: 30 min.

Frequently Asked Questions (FAQs)

Can I elute my plasmid from PureLink columns with water instead of TE?

For any silica columns, elution with water is generally possible. However, a buffer is preferred for stability and accuracy of absorbance readings, as pure water can have a very low pH (4 - 5).

What are endotoxins and why are their levels important for transfection?

Endotoxins are typically any cell-associated bacterial toxins that are part of the outer surface of the cell wall of gram-negative bacteria. Endotoxins can influence cell growth, cell differentiation, contractility, and protein expression in mammalian cells. Endotoxins are released during bacterial lysis, and they can subsequently reduce transfection efficiency and protein expression levels. Please review the following article for more information about endotoxins: Butash KA et al. (2000) Reexamination of the effect of endotoxin on cell proliferation and transfection efficiency. Biotechniques 29(3): 610-614, 616, 618-619.

What is the best way to determine the quality/purity of my plasmid DNA?

The ratio of absorbance at 260 nm to the absorbance at 280 nm (A260/A280) is typically used to measure purity of the sample. For DNA, the ideal A260/A280 ratio is 1.8, but it can be in the range of 1.7 - 1.9. The A260/A230 ratio is also used to determine if contamination is present. For DNA, the ideal A260/A230 ratio is between 1.8 and 2.0. DNA purity can also be examined by gel analysis. For plasmid DNA, look for a strong, single band (perhaps with a few extra bands representing multimers of the desired molecule). For genomic DNA, look for high average fragment sizes.

Do you offer a kit for plasmid extraction with endotoxin reduction?

We offer the following silica column-based plasmid purification kits for isolation of endotoxin-free or low-endotoxin plasmid DNA:
- PureLink Expi Endotoxin-Free Maxi Plasmid Purification Kit (Cat. No. A31217, A31231, A33073) (<0.1 EU/µg endotoxins):
https://www.thermofisher.com/order/catalog/product/A31217
- PureLink Fast Low-Endotoxin Midi (Cat. No. A35892, A36227) or Maxi (Cat. No. A36228, A35895) Plasmid Purification Kits (<1 EU/µg endotoxins):
https://www.thermofisher.com/order/catalog/product/A35892
https://www.thermofisher.com/order/catalog/product/A36228

Will kit performance be affected if I exceed culture input volumes with PureLink Fast Low-Endotoxin Plasmid Purification kits?

Our recommendation is to not exceed the maximum recommended culture volumes specified in the kit manuals; using larger culture volumes means increased cell mass, which will result in reduced plasmid yield and purity.

Can I isolate low-copy plasmids with PureLink Fast Low-Endotoxin Plasmid Purification kits?

We recommend using high-copy number plasmids cultured in LB medium to obtain maximal yield and purity of plasmid DNA. You can use low-copy plasmids, but please be aware that the expected yield and purity will be considerably lower.

Can I use enriched media (i.e., Terrific broth) to grow the overnight culture when using PureLink Fast Low-Endotoxin Plasmid Purification kits?

Avoid using rich media for bacterial cultures. If using a rich medium is necessary, ensure that the recommended cell mass is not exceeded by optimizing the growth time so that the OD600 stays at <4.0, or by processing a smaller culture volume (<100 mL).

What is the recommended OD600 range for the overnight culture when using PureLink Fast Low-Endotoxin Plasmid Purification kits?

We recommend using bacterial cultures at OD600 of 2.0 - 4.0.

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