Can the isolated DNA be manipulated by means other than PCR – such as restriction digestion or (more importantly) PCR sequencing?
The DNA prepared with Extract-N-Amp is not adequately purified for most downstream applications – in fact, it isn’t purified at all. PCR is so sensitive that very little DNA, and hence, very little of the accompanying impurities are added to the reaction. The amount of DNA generally used for restriction digests and visualization on ethidium bromide-stained gels will likely contain enough impurities to inhibit a restriction enzyme. On the other hand, we have obtained good sequencing data from PCR products generated with the Extract-N-Amp Blood Kit.
Related Questions
- Can the isolated DNA be manipulated by means other than PCR – such as restriction digestion or (more importantly) PCR sequencing?
- What is the DNA sequence around the left border of pROK2 and where do the PCR isolation and DNA sequencing primers anneal?
- Can the isolated DNA be used in applications other than PCR – such as restriction digestion or Southern blots?
