NA0100

Sigma-Aldrich

PhasePrep BAC DNA Kit

Scalable method for isolating large-molecular weight plasmids

Synonym(s):
BAC Kit
NACRES:
NA.52
Pricing and availability is not currently available.

Quality Level

storage temp.

15-25°C

Related Categories

General description

The PhasePrep BAC DNA Kit offers a scalable and cost-effective method for isolating large-molecular weight plasmids such as Bacterial Artificial Chromosomes (BAC) from recombinant E. coli cultures. The same kit can be used for preparations of four different sizes; sufficient reagents are provided for 300 micro, 180 mini, 30 midi, or 15 maxi preparations. Up to 2,12, 50, or 100 mg of BAC DNA can be recovered from 5, 40, 250, or 500 ml of overnight recombinant E. coli culture, respectively. The purified BAC DNA contains very low levels of endotoxin (£10 EU/mg DNA).

Recombinant E. coli culture is harvested by centrifugation and subjected to a modified alkaline-SDS lysis procedure. Nucleic acid is precipitated from the cleared lysate; residual RNA is removed by a short digestion at elevated temperature with an RNase cocktail. Endotoxins and other impurities are removed by simple temperature-mediated extraction and phase separation. Finally the BAC DNA is selectively precipitated from solution.
The PhasePrep BAC DNA Kit offers a scalable and cost-effective method for isolating large-molecular weight plasmids such as Bacterial Artificial Chromosomes (BAC) from recombinant E. coli cultures. The same kit can be used for preparations of four different sizes; sufficient reagents are provided for 300 micro, 180 mini, 30 midi, or 15 maxi preparations. Up to 2,12, 50, or 100 mg of BAC DNA can be recovered from 5, 40, 250, or 500 mL of overnight recombinant E. coli culture, respectively. The purified BAC DNA contains very low levels of endotoxin (≤10 EU/μg DNA).

Recombinant E. coli culture is harvested by centrifugation and subjected to a modified alkaline-SDS lysis procedure. Nucleic acid is precipitated from the cleared lysate; residual RNA is removed by a short digestion at elevated temperature with an RNase cocktail. Endotoxins and other impurities are removed by simple temperature-mediated extraction and phase separation. Finally the BAC DNA is selectively precipitated from solution.

The recovered BAC DNA is predominantly in its super-coiled form, free of RNA contamination, and ready for immediate use in downstream applications, such as sequencing, restriction digestion, cloning, and PCR.

Application

The recovered BAC DNA is predominantly in its super-coiled form, free of RNA contamination, and ready for immediate use in downstream applications, such as sequencing, restriction digestion, cloning, and PCR.
PhasePrep BAC DNA Kit has been used to isolate bacterial artificial chromosomes.

Features and Benefits

  • Typical DNA yields of 2-100 μg from 5-500 mL of overnight cultures
  • No phenol/chloroform extraction required
  • Allows possible micro to maxi preps with the same kit
  • No long waits for drip columns

Other Notes

The PhasePrep BAC DNA Kit offers a highly scalable, rapid, cost-effective method for isolating high molecular weight plasmids such as Bacterial Artificial Chromosomes (BACs) from recombinant E. coli cultures.
For additional information, please see www.sigma-aldrich.com/genelutehp.

Legal Information

PhasePrep is a trademark of Sigma-Aldrich Co. LLC

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Precautionary Statements

Target Organs

Central nervous system

RIDADR

UN 3316 9

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis
Certificate of Origin
Structure, phylogeny, allelic haplotypes and expression of sucrose transporter gene families in Saccharum
Qing Zhang
BMC Genomics, 17 (2016)
Evolution and expression of the fructokinase gene family in Saccharum
Yihong Chen
BMC Genomics, 18, 197-197 (2017)
Comparative structural analysis of Bru1 region homeologs in Saccharum spontaneum and S. officinarum
Jisen Zhang
BMC Genomics, 17 (2016)
S S Kwek et al.
Oncogene, 28(17), 1892-1903 (2009-03-31)
Co-amplification at chromosomes 8p11-8p12 and 11q12-11q14 occurs often in breast tumors, suggesting possible cooperation between genes in these regions in oncogenesis. We used high-resolution array comparative genomic hybridization (array CGH) to map the minimal amplified regions. The 8p and 11q...

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