Hi,
Anybody knows how does circular, uncut pGEM-T (with fragment of course) migrate on agarose? How many bands does it yield at at what level of the Ladder?
Thx
0
2 replies to this topic
#2
bob1
-
- Global Moderators
-
- 4,350 posts
Thelymitra pulchella
224
Excellent
Excellent
Posted 31 August 2011 - 06:02 PM
Assuming there is no linear or supercoiled DNA then it should result in 1 band only. If you just have a standard plasmid extraction you may see 3 bands depending on the integrity of the DNA you have extracted. These bands will be the supercoiled, the nicked (i.e. circular, single strand breakage) and the linear (double strand break). If you see more bands then there has been further digestion of the DNA, or you have degraded genomic DNA or RNA in your prep.
#3
pDNA
-
- Active Members
-
- 489 posts
Veteran
14
Good
Good
Posted 01 September 2011 - 12:03 PM
...or if isolated from a recA positive strain you'll see plasmid multimers (two/three/four/etc-times the vector size).
Regards,
p
Regards,
p
Also tagged with one or more of these keywords: plasmids, pGEM-T
Protocols and Techniques Forums →
Molecular Cloning →
|
|
|
||
Protocols and Techniques Forums →
Molecular Biology →
Time on ice before preppingStarted by Guest_hogthehedge_* , 14 Jul 2012  E coli, LB, Maxipreps, Plasmids and 1 more...
|
|
|
||
Protocols and Techniques Forums →
Molecular Cloning →
Easy to purify Plasmid using Lambda genesStarted by Guest_Koeng_* , 05 Jul 2012 Microbiology, plasmids
|
|
|
||
Protocols and Techniques Forums →
Microbiology →
|
|
|
||
Protocols and Techniques Forums →
Molecular Biology →
eGFP vs eGFP-N1Started by Guest_sabalab_* , 25 May 2012 eGFP, plasmids, cloning and 1 more...
|
|
|
