Catalog

18000

BAC DNA MiniPrep Kit

( 25 preps )
Norgen Biotek

172.06$

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Additional description

BAC DNA MiniPrep Kit (Cat# 18000 )

For the rapid preparation of BAC DNA

  • High DNA recovery
  • Alcohol-free protocol
  • Convenient spin column protocol

Product Description

Norgen’s BAC DNA MiniPrep Kit is designed for the rapid preparation of Bacterial Artificial Chromosomes (BACs)and other large DNA constructs from small batch cultures of Escherichia coli.The BAC DNA MiniPrep Kit typically recovers between 0.6 and 1 µg of DNA from 3 mL of bacterial culture.Purification of constructs up to 130 kbp in size have been verified.

Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. Briefly, thebacteriaharbouring the plasmidis pelleted and lysed, and the released plasmid DNA isbound to Norgen's column in the presence of Binding Solution (BIND).Under these conditions only the DNA will bindto Norgen's resin whilemost ofthe contaminating RNAand cellular proteinaceous componentsare removed in the flowthrough. The bound DNAis then washed to remove anyremaining impurities(WASH).Lastly, the purified DNA is eluted into30 -50µL of the provided ElutionBuffer or water(ELUTE).Pleaseseeprocedure flowchart to the right.

Norgen's BAC DNA Miniprep Kit purifies DNA without alcohols, a significant improvement over other techniques to achieve full compatibility with alcohol-sensitive applications.The purified DNA is free of genomic DNA, RNA and cellular proteins and is suitable for many downstream applications.

FEATURES AND BENEFITS

  • High Recovery -0.6 to 1 µg of BAC DNA can be purified from 3.0 mL of bacterial culture.
  • Alcohol-free protocol -No alcohols are used durig the purification process, thus the DNA is compatible with alcohol sensitive downstream applications.
  • Fast and easy processing - Rapid spin-column format allows for the processing of multiple samples in 1 hour.
  • Recovered DNA is suitable for downstream applications - Purified BAC DNA is fully compatible with restriction enzyme digestions and sequencing.
  • High binding capacity of columns -The binding capacity of the columns in the BAC DNA MiniPrep Kit is 25 µg

Specifications

Purified BACDNAis ready for a range of downstream applications including:

  • sequencing
  • transformation
  • restriction digestion

Figure 1.Higher DNA Yield than Competitor. BAC DNA from 3 mL overnight cultures was isolated with the Norgen BAC MiniPrep Kit (Lanes 1-3) and with a competitor’s kit (Lanes 4-6).Ten microliters of 50 µL of recovered DNA was loaded in each lane of a 1X TAE, 0.9% agarose gel.Lane M is Norgen’s UltraRanger 1kb DNA Ladder.Note the quality, consistency and yield of the DNA isolated using Norgen’s kit (Lanes 1-3).

Figure 2. Full Compatibility with Digests. BAC DNA isolated with Norgen’s BAC DNA MiniPrep Kit is fully digestible.In Figure 2, 10 µL of two different isolated BACs (50 µL elution) were digested with 2 units of HindIII (Lanes 3 and 4) in a 20 µL reaction at 37oC overnight.Lanes 1 and 2contain 10 µL of uncut plasmid, and Lane M is Norgen’s UltraRanger 1kb DNA Ladder.All DNA was resolved on a 1X TAE, 0.9% agarose gel.

APPLICATIONS

Kit Specifications
Column Binding Capacity
25 µg
Average Yield
0.6 - 1 µg
Size of Plasmids Purified
Up to 130 kbp
Time to Complete 10 Purifications
1 hour


BAC DNA MiniPrep Kit Contents

1. Resuspension Buffer
2. Lysis Solution
3. Neutralizaation Solution
4. Binding Solution
5. RNAse A
6. RNase T1
7.Wash Solution
8. Elution Buffer
9. Micro Spin Columns
10.Collection Tubes
11. Elution Tubes
12. Product Insert


Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. The RNAse A and RNAse T1 should be stored at -20°C upon arrival. Once RNAse A has been added to the Resuspension Buffer, however, the solution should be stored at 4oC. All the reagents should remain stable for at least 1 year in their unopened containers.