69281
DNase Shotgun® Cleavage Kit
Random, double-stranded cleavage of any DNA for shotgun cloning
For general questions please contact our Customer Service:
Merck KGaA
Frankfurter Str. 250
64293 Darmstadt
Germany
Phone: +49 6151 72-0
Fax: +49 6151 72 2000
15 February 2012
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The DNase Shotgun® Cleavage Kit is designed
to generate random DNA fragments from
any DNA sample in preparation for cloning.
The sample DNA is cleared with DNase I in
the presence of Mn2+, which causes random
double-stranded cleavage of the DNA
molecule (Anderson 1981). The DNase I in the
kit is specially prepared in a buffer lacking
Mg2+ to prevent single-strand nicking.
Fragments of almost any size can be generated by adjusting the amount of enzyme and/or time of reaction. Specific size ranges can be isolated by agarose gel electrophoresis, and the ends repaired using a DNA polymerase (e.g., to generate blunt ends treat with T4 DNA polymerase in the presence of dNTPs). The Single dA Trailing Kit repairs DNA ends and adds a 3′-dA residue. The processed DNA fragments can be easily cloned into any vector with single 3′-dT or 3′-dU overhangs, such as one of the Novagen AccepTor™ Vectors. Advantages of this cloning strategy include: prevention of tandem inserts, low nonrecombinant background, and no requirements for special linkers or additional fractionation steps. This patented approach is used to generate protein domain expression libraries with the Novagen NovaTope® System.
Fragments of almost any size can be generated by adjusting the amount of enzyme and/or time of reaction. Specific size ranges can be isolated by agarose gel electrophoresis, and the ends repaired using a DNA polymerase (e.g., to generate blunt ends treat with T4 DNA polymerase in the presence of dNTPs). The Single dA Trailing Kit repairs DNA ends and adds a 3′-dA residue. The processed DNA fragments can be easily cloned into any vector with single 3′-dT or 3′-dU overhangs, such as one of the Novagen AccepTor™ Vectors. Advantages of this cloning strategy include: prevention of tandem inserts, low nonrecombinant background, and no requirements for special linkers or additional fractionation steps. This patented approach is used to generate protein domain expression libraries with the Novagen NovaTope® System.
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| Avoid freeze/thaw | Yes | |||||||||||||||||
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| Storage | -20°C | ||
| Ship |
Shipped with Blue Ice or with Dry Ice
Multiple Toxicity Values, refer to MSDS |
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