DNA Sequencing / Cloning ?
Posted 20 August 2010 - 04:12 PM
I would just like to ask about DNA sequencing methods; for my dissertation, i spent months extracting DNA and then cloning my sequence using bacterial cultures to produce a sample that we were able to sequence from. Perhaps it was the rush of handing the project in on time that made me blank but could someone possiably tell me why DNA from a bacterial cell was sequenceable whereas it wasn't from the original mammalian cell? I guess my point is, what was the purpose of the cloning step and does sequencing via bacteria produce more accurate results from that of a mammals?
I'm guessing that perhaps the genome is smaller thus makes it more accurate to sequence from?
I would greatly appreciate some wisdom and also are there any other methods for sequencing DNA other than cloning the sequence first?
Many thanks from a fledgling scientist!
Posted 20 August 2010 - 06:05 PM
It's not that you can't sequence DNA recovered from mammalian cells -- you can (it's the same DNA, after all), it's just so much easier to recover sufficient quantities of your gene in pure form from bacterial cells than it would be to recover an equal number in as pure form from mammalian cells.
Posted 21 August 2010 - 04:32 AM
Posted 21 August 2010 - 12:31 PM
Posted 31 August 2010 - 06:08 PM