DNA & RNA Purification

PhasePrep™ BAC DNA Kit

Description: 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.

Procedure: The kit utilizes a phase separation procedure that can be scaled to micro, mini, midi, or maxi prep sizes. Sufficient reagents are provided for 300 micro, 180 mini, 30 midi, or 15 maxi preps.

Application: The purified BAC DNA contains very low levels of endotoxins and is ready for immediate use in Automated Sequencing (Figure 1), PCR, Restriction Enzyme Digestion (Figure 2), Cloning and other common applications.
GenElute™ PhasePrep™ BAC DNA Flow Chart

Beneficial Features

Starting Material 5, 40, 250 or 500 ml overnight culture for micro, mini, midi or maxipreps, respectively
Yield Typical DNA yields of 2 - 100 µg from 5 - 500 ml of overnight cultures
Speed Less than 1 hour; no tedious gravity-flow columns
Safe No phenol/chloroform extraction required
Versatile Allows possible micro to maxi preps with the same kit

Order Information

Product No. Name Culture Volume Expected Yield Technical Manual .pdf
NA0100 PhasePrep BAC DNA Kit 5 - 500 ml 2 - 100 µg X

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Sample Data

High-quality BAC DNA for Automated Sequencing
Figure 1. BAC DNA was purified with the PhasePrep BAC DNA Kit and sequenced with a custom primer using BigDye™ Terminator Chemistry. Sequencing reactions were resolved on an ABI 3700 sequencing instrument.

Data provided by the Genome Sequencing Center at Washington University, St. Louis
Click Image to View Sequence in its Entirety
Fig. 1. Click to enlarge.

Purified BAC DNA is suitable for restriction digestion
Figure 2. Restriction digestion of BAC DNA isolated with the PhasePrep BAC DNA Kit and two other suppliers (C and Q). BAC DNA samples were purified from overnight cultures of E. coli HB101b transformed with a pbelloBAC11 clone. Approximately 1 µg of DNA from each sample was digested with 20 units of Eco R V at 37°C for 4 hours. One half of each digestion (approximately 0.5 micrograms) was separated by overnight electrophoresis in a 1% agarose gel. Lanes 1 to 8 were BAC DNA purified with Sigma kit, lane 9 was BAC DNA purified with Supplier C's kit, and lane 10 was BAC DNA purified with Supplier Q's kit. The molecular marker used at the first and last lanes was a 1kb DNA Ladder (M) (Product No. D0428).

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