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Home Q&AcWhy can¡¦t the eluted plasmid be cut by restriction enzymes properly?

Plasmid DNA Extraction Kit(PD004, PD100, PD300) --Q&A
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Q Can the plasmid DNA extraction Kit be applied on Gram (+) bacteria?
Q Why do we get eluted plasmid contaminated with RNA?
Q Why can¡¦t the eluted plasmid be cut by restriction enzymes properly?
A It is possible that salt residue in buffers or ethanol residue in the Wash Buffer is not removed completely, thus affecting the downstream reaction. In case of salt residue, wash the column twice with Wash Buffer. In case of ethanol residue, after washing with Wash Buffer, make sure that the flow-through is discarded and centrifuge the column at full speed for 3 minutes. If necessary, centrifuge for a few minutes more to ensure complete removal of ethanol.
Another reason is that the plasmid is denatured. Denaturation occurs if incubation in PD2 Buffer is too long. This can be seen during electrophoresis. After PD2 Buffer is added, DO NOT incubate for more than 5 minutes.
Q Why does the Plasmid DNA Extraction Kit require two washings with two types of wash buffers (W1 and Wash Buffer) while competitors kits require one washing?
Q Eluted plasmid appeared degraded after storage for an extended period of time.
Q Why was Restriction enzyme digestion unsuccessful?

Genomic DNA Extraction Kit(GT004,GT050,GT300,GB004,GB100,GB300,GP004,GP100)--Q&A
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Q Why doesn¡¦t the Eluted DNA perform well in downstream applications?
Q Why does the genomic DNA band appear smeared in agarose gel electrophoresis?
Q What is the key to successfully isolating genomic DNA of good yield and quality?
Q Why does the sample appear viscous after sample Lysis and not pass through the column easily, even when less than the suggested maximum sample amount is used?
Q When Buffy coat is used, the column tends to be clogged.

Gel/PCR DNA Fragments Extraction Kit (DF004, DF100, DF300)--Q&A

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