BAC DNA purification—NucleoBond Xtra BAC

Purify large DNA fragments such as cosmids, bacteriophage P1 clones, PACs, and BACs using anion exchange technology

NucleoBond Xtra BAC provides second-generation anion exchange for large-construct plasmid DNA. The high selectivity and specific characteristics of the optimized silica matrix make this anion exchange kit ideal for efficiently purifying very large constructs, such as P1, BACs, and PACs. For example, using the NucleoBond Xtra BAC kit, it takes just 75 minutes to isolate BAC DNA free of all types of RNA, proteins, metabolites, dyes, and carbohydrates.

LyseControl, a blue pH indicator reagent present in the lysis buffer used in the NucleoBond Xtra BAC procedure, ensures complete and efficient alkaline lysis, providing optimal yields. Upon addition of lysis buffer to the resuspended cells, the cell suspension turns blue. To avoid contamination with genomic DNA, the blue solution is gently inverted up to five times until a homogeneous and 100% colorless solution is achieved. After a five-minute incubation, neutralization buffer is added and LyseControl turns colorless. Traces of blue LyseControl indicate insufficient mixing. Using LyseControl ensures sufficient mixing and complete neutralization, which are necessary to renature plasmid DNA, precipitate proteins/gDNA, and remove SDS, which inhibits plasmid binding.

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Cat. #	Product	Size	Price
740436.25	NucleoBond® Xtra BAC	25 Preps	USD $785.00
NucleoBond Xtra BAC (25) 25 preps for the isolation of low-copy larg construct plasmid DNA - NucleoBond Xtra BAC Columns, buffers, RNase A Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents You May Also Like Image Data Resources Back NucleoBond Xtra BAC is suitable for restriction digestion: BAC DNA (300 kb) was isolated in triplicate from 500 ml E. coli DH5a using NucleoBond Xtra BAC and competitor products (Q and I) *NucleoBond Xtra BAC is suitable for restriction digestion: BAC DNA (300 kb) was isolated in triplicate from 500 ml E. coli* DH5a using NucleoBond Xtra BAC and competitor products (Q and I).** After precipitation, BAC DNA was reconstituted in 1,000 μl TE buffer and 5 μl of each sample was used for analysis on a 1% TAE agarose gel. Obtained yields: MN: 150 μg, Q: 44 μg, I: 75 μg. Restriction digestion of BAC DNA isolated with NucleoBond Xtra BAC. BAC DNA samples were purified from overnight cultures of E. coli DH5a transformed with a pBAC10 clone. Approximately 3 μg of DNA from each sample was digested with 3 units of MspI, HindIII, or EcoRI at 37°C for 2 hr. Digestions were analyzed on a 1% TAE agarose gel. Lane 1: undigested BAC DNA, lane 2: MspI-digested, lane 3: HindIII-digested, lane 4: EcoRI-digested, lane M: marker. Back High selectivity and specific characteristics of the optimal silica matrix make this anion exchanger ideal for the purification of very large constructs, such as P1, BACs, and PACs High selectivity and specific characteristics of the optimal silica matrix make this anion exchanger ideal for the purification of very large constructs, such as P1, BACs, and PACs. Using the NucleoBond Xtra BAC kit, it takes just 75 min to isolate, for example, BAC DNA free from any kind of RNA, proteins, metabolites, dyes, and carbohydrates. Low amounts of impurities, which often act as strong inhibitors for enzymatic processing, transfections, transcriptions, etc., are completely removed. Back Highest yield in less time – comparison to competitor kits Highest yield in less time – comparison to competitor kits. BAC DNA (300 kbp) was isolated in triplicate from 500 ml E. coli DH5a using NucleoBond Xtra BAC and competitor products (Q and I). After precipitation, BAC DNA was reconstituted in 1000 μl TE buffer and 5 μl of each sample was used for analysis on a 1 % TAE agarose gel. Obtained yields: MN: 150 μg, Q: 44 μg, I: 75 μg. Back 740436.25: NucleoBond Xtra BAC
740436.10	NucleoBond® Xtra BAC	10 Preps	USD $343.00
NucleoBond Xtra BAC (10) 10 preps for the isolation of low-copy large construct plasmid DNA - NucleoBond Xtra BAC Columns, buffers, RNase Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents You May Also Like Image Data Resources Back 740436.10: NucleoBond Xtra BAC Back NucleoBond Xtra BAC is suitable for restriction digestion: BAC DNA (300 kb) was isolated in triplicate from 500 ml E. coli DH5a using NucleoBond Xtra BAC and competitor products (Q and I) *NucleoBond Xtra BAC is suitable for restriction digestion: BAC DNA (300 kb) was isolated in triplicate from 500 ml E. coli* DH5a using NucleoBond Xtra BAC and competitor products (Q and I).** After precipitation, BAC DNA was reconstituted in 1,000 μl TE buffer and 5 μl of each sample was used for analysis on a 1% TAE agarose gel. Obtained yields: MN: 150 μg, Q: 44 μg, I: 75 μg. Restriction digestion of BAC DNA isolated with NucleoBond Xtra BAC. BAC DNA samples were purified from overnight cultures of E. coli DH5a transformed with a pBAC10 clone. Approximately 3 μg of DNA from each sample was digested with 3 units of MspI, HindIII, or EcoRI at 37°C for 2 hr. Digestions were analyzed on a 1% TAE agarose gel. Lane 1: undigested BAC DNA, lane 2: MspI-digested, lane 3: HindIII-digested, lane 4: EcoRI-digested, lane M: marker. Back High selectivity and specific characteristics of the optimal silica matrix make this anion exchanger ideal for the purification of very large constructs, such as P1, BACs, and PACs High selectivity and specific characteristics of the optimal silica matrix make this anion exchanger ideal for the purification of very large constructs, such as P1, BACs, and PACs. Using the NucleoBond Xtra BAC kit, it takes just 75 min to isolate, for example, BAC DNA free from any kind of RNA, proteins, metabolites, dyes, and carbohydrates. Low amounts of impurities, which often act as strong inhibitors for enzymatic processing, transfections, transcriptions, etc., are completely removed. Back Highest yield in less time – comparison to competitor kits Highest yield in less time – comparison to competitor kits. BAC DNA (300 kbp) was isolated in triplicate from 500 ml E. coli DH5a using NucleoBond Xtra BAC and competitor products (Q and I). After precipitation, BAC DNA was reconstituted in 1000 μl TE buffer and 5 μl of each sample was used for analysis on a 1 % TAE agarose gel. Obtained yields: MN: 150 μg, Q: 44 μg, I: 75 μg.

Overview

Optimized column design—complete one prep in approximately 75 minutes
Column filter included—allows parallel lysate clearing and loading onto the column
Optimized silica matrix—yields up to 150 μg of purified large-construct plasmid DNA
Transfection-grade plasmid DNA—obtained using proven anion-exchange technology
LyseControl—visually confirms completion of lysis/neutralization, maximizing yield

More Information

Technology	Anion exchange
Format	Gravity-flow columns
Lysate clarification	Column filters
Starting material	250–750 ml
Vector size	<300 kb
Typical yield	10–100 µg
A_260/A280	1.80–1.95
Isopropanol precipitation	Centrifugation
Preparation time	75 min/prep
Binding capacity	150 µg

Applications

Purification of P1, BACs, PACs, and other large constructs from E. coli
Typical downstream applications: cloning, PCR, restriction analysis, sequencing, and transfection

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.

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BAC DNA purification—NucleoBond Xtra BAC

Notice to purchaser

NucleoBond Xtra BAC is suitable for restriction digestion: BAC DNA (300 kb) was isolated in triplicate from 500 ml E. coli DH5a using NucleoBond Xtra BAC and competitor products (Q and I)

High selectivity and specific characteristics of the optimal silica matrix make this anion exchanger ideal for the purification of very large constructs, such as P1, BACs, and PACs

Highest yield in less time – comparison to competitor kits

740436.25: NucleoBond Xtra BAC