plasmid restriction mapping query

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aditya5
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plasmid restriction mapping query

Post by aditya5 » Fri Sep 10, 2010 3:12 am

Hi All,

We have performed plasmid isolation using miniprep isolation kit and done restriction mapping of the said plasmid. On running unrestricted plasmid on the agarose gel electrophoresis we got 2 prominent bands and some other high mol wt bands. our plasmid is about 2.5 kbps one of the prominent band was 2.5 kbps but other was about 7 - 8 kbps and other light bands were higher than that. we performed restriction digestion using bamHi and got single band but at aroubd 3.5 Kbps. what is the reason for this. if the plasmid size is 2.5 kbps why on restriction size is more and what is this high 7-8 kbps band.

i am a microbiologist and new to this molecular biology trials. So the question may seem silly. would appreciate any guidance. :?:

thanks in anticipation


aditya

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JackBean
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Post by JackBean » Fri Sep 10, 2010 6:23 am

regarding uncut plasmid
http://en.wikipedia.org/wiki/Plasmid#Conformations

cut plasmid:
hard to say, did you read correctly the ladder? Didn't the plasmid contain some gene or something?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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