Login

Join for Free!
112474 members


I would like to prepare DNA ladder.Who has a protocol?

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderator: BioTeam

I would like to prepare DNA ladder.Who has a protocol?

Postby arm_chair27 » Thu Jun 07, 2007 3:45 pm

I would like protocol to prepare DNA ladder. Someone help me!
arm_chair27
Garter
Garter
 
Posts: 3
Joined: Mon Mar 26, 2007 12:46 pm

Postby wbla3335 » Thu Jun 07, 2007 5:41 pm

We used to make our own just by cutting a plasmid with some enzymes. I can't remember which plasmid we used or which enzymes. Just decide what range of sizes you want, then sit down with a programme that can show you fragment sizes, and find a good combination of enzymes/plasmid that will give you what you want.
wbla3335
Coral
Coral
 
Posts: 227
Joined: Thu Aug 17, 2006 7:20 am

Postby blcr11 » Fri Jun 08, 2007 7:46 am

Lambda phage DNA digested with HindIII is what everybody used to use. I think maybe a PstI digest of lambda DNA is also a possibility. That means you have to prepare or buy some lambda DNA and one or more restriction enzymes, but if you can afford to buy the DNA and enzymes, you are advised to purchase some ready-made (or ready-cut) marker DNAs. Much simpler and probably more reliable. If you only need sizes up to 3-4 kbp, there is probably of digest of pBR322 that would work, though without looking at a map I don't know what enzyme to suggest.
blcr11
Viper
Viper
 
Posts: 672
Joined: Fri Mar 30, 2007 4:23 am



Return to Molecular Biology

Who is online

Users browsing this forum: No registered users and 1 guest

cron