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This is fast and efficient method for site directed mutagenesis using Type IIS restriction enzymes. Here this method would be applied to mutate the BSMBI cut site within the hygromycine resistant gene of V0200 mammalian vector.
DNA Purification Using the GenElute™ PCR Clean-Up Kit for rapid purification of double-stranded PCR amplification products.
Incubate the tubes at 80°C for 10 minutes and place the tube and heat block out of the heater on the bench cool down the digested parts gradually in room temperature. Bacterial transformation of each ligation reaction into DH5?-T , fast protocol: incubate DNA and cells on ice for 30 minutes and then spread on AMP agar plates and incubate in 37°C overnight. Array-based comparative genomic hybridization (aCGH) is a powerful tool for detecting gene copy number gains and losses associated with chromosome abnormalities.

Enzo’s proprietary labeling technology generates consistent results that exceed industry standards, increasing accuracy of variant detection, minimizing manual data analysis, increasing efficiency, and reducing overall sample analysis time. Enzo’s proprietary labeling technology delivers excellent DNA yields with superior dye incorporation leading to the highest specific activity of labeling.
Enzo’s CGH protocol requires no DNA digestion or restriction enzymes, saving time and preventing DNA loss during restriction enzyme cleanup. Enzo’s CYTAG™ TotalCGH Labeling kit includes all the components needed to perform CGH+SNP analysis. Quick, easy kit for purifying DNA from PCR, gels and labeling reactions for a variety of downstream applications.
With designing a pair of mutagenesis primers that the mutated sequence is near and downstream of the recognition site of the type IIs restriction enzyme.

Detecting chromosomal aberrations by aCGH is faster, more robust and provides superior results over other technologies such as FISH and G-banding karyotyping, thus providing a greater understanding of the role of chromosomal changes in genetic diseases and cancers. Enzo’s proprietary labeling technology generates the highest specific activity of labeling.
DNA was labeled with the Enzo’s CYTAG™ TotalCGH Labeling Kit (left) or a competitor’s kit (right). The Enzo kit cleanly identifies the sequence at single nucleotide polymorphisms (SNPs) while quantification of the SNP analysis of DNA labeled with a competitor’s labeling system is not as clean.

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  2. 125:
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