M13 for cloning? - (May/19/2008 )
Back in the early '90s I did a lot of cloning and sequencing using M13. I'm now planning a project that will involve lots of cloning and sequencing, but when I look at the cost of all these fancy new cloning kits, and I do the math -- well, it got me to thinking about using M13 again. Some of these kits are nice, though, saving a lot of work. A quick search through the literature shows that M13 for cloning isn't used much anymore. Is there some middle road I don't know about, or is everyone richer than they used to be?
It does depend upon what kind of cloning and what downstream application you are looking for. But for all practical purposes I think M13 is out and for a good reason. You don't need kits for everything, and yet you could do better cloning than using M13 methods.
It would help if you tell us what exactly you want to do. Some methods here..
TOPO
PCR and TA cloning
PCR and sequencing
Recombineering
M13 was popular when single stranded DNA was required for sequencing. With Sanger dideoxy sequencing, double stranded DNA is fine, and the utility of M13 is very limited.
You could also just do your sequencing from whatever plasmid you've cloned into. Are you planning on doing anything else with your clones, like expression? As long as you have the correct sequencing primers for the plasmid you've selected for your work, you should be laughing.
Hallo, i think kits for sequencing is best way since price of work, media, chemistry and mainlly cost of the time will be not equal. Additionaly M13 is suitable for short fragment sequencing, 500bp. Our staf can 1-1.2 kb
good luck