BAC purification—NucleoBond BAC 100
The NucleoBond BAC 100 Kit is designed for purifying large DNA fragments such as cosmids, bacteriophage P1 clones, P1-derived artificial chromosomes (PACs), and bacterial artificial chromosomes (BACs), without phenol/chloroform extraction. The procedure takes less than one hour to isolate BAC DNA free from any kind of RNA, proteins, metabolites, dyes, and carbohydrates. After purification, all nucleic acids show absorbance ratios of A260/A280 as high as 1.8–2.0.
The NucleoBond BAC 100 Kit is designed for purifying large DNA fragments such as cosmids, bacteriophage P1 clones, P1-derived artificial chromosomes (PACs), and bacterial artificial chromosomes (BACs), without phenol/chloroform extraction. The procedure takes less than one hour to isolate BAC DNA free from of RNA, proteins, metabolites, dyes, and carbohydrates. After purification, all nucleic acids show absorbance ratios of A260/A280 as high as 1.8–2.0.
Low amounts of impurities, which often act as strong inhibitors for enzymatic processing, sequencing, transfections, transcriptions, etc., are removed completely.
NucleoBond Folded Filters, which are specially designed to eliminate the centrifugation step after the alkaline lysis step, are provided with the kit. Using the filters reduces hands-on time and eliminates plastic waste common to other filtration systems. In approximately 10 minutes of unattended operation, complete removal of SDS and cellular debris from plasmid samples is achieved. NucleoBond Folded Filters do not induce shearing of large DNA constructs, such as PACs or BACs.
The patented anion-exchange resin in the NucleoBond column is known to provide high-purity DNA equivalent or superior to that obtained by two successive rounds of CsCl ultracentrifugation. The plasmid purification procedure is performed in the absence of toxic material such as phenol, chloroform, ethidium bromide, or CsCl.
Overview
- For purification of P1 constructs, BACs, PACs, and other large constructs
- Vectors up to 300 kb can be purified
- Increased buffer volumes included
- NucleoBond Folded Filters included for lysate clarification
- Anion-exchange technology
More Information
| Technology | Anion exchange |
| Format | Maxi |
| Lysate clarification | Folded filters |
| Starting material | 100–500 ml |
| Vector size | <300 kb |
| Typical yield | 10–100 µg |
| A260/280 | 1.80–1.95 |
| Preparation time | 120 min/2–4 preps |
Applications
- Cloning
- Sequncing
- PCR
- Transformation
- Restriction analysis
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
Tip for a successful transfection—remove endotoxins!
Endotoxins are copurified during plasmid preparations from bacterial lysates and affect eukaryotic cell survival, ultimately leading to a lower transfection efficiency. Innovative mini-format technology reduces endotoxins in your plasmid prep, leads to more successful transfections, and simplifies your workflow.
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