Principle steps from dna extraction to pcr to sequencing?

Depending on where your template DNA came from, you shouldn't need to do much to it before PCR. If the primers are designed correctly (i.e. they won't hybridize to unwanted targets), only your sequence of interest will be amplified. Quantification can be performed in a variety of ways, including checking the band intensity (if your template is of known size) on a gel. In the PCR reaction, many more copies of primers are needed than copies of template. Following PCR amplification, run a gel to confirm presence and proper size of the product, and then cut that product out of the gel to purify it from the other components of the PCR reaction. uming there is enough product, the resulting DNA can be sent to be sequenced without cloning (but this may be necessary for future use depending on your needs). To fill in any gaps in the protocol, or to answer any additional questions, I recommend looking at Qiagen's kits - they're the most commonly used in the lab. To answer your last question, yes, RNA can be purified from a gel in a similar manner, but it is much less stable than DNA, so it is more difficult to purify without losing a significant amount of product in the process.