Search found 33 matches

by genetherapy
Mon Jul 30, 2012 11:25 am
Forum: Molecular Biology
Topic: How to calculate mass of target Dna in ng
Replies: 1
Views: 3121

Look at this web-sites,the informations that they have will help you

http://www.molecularstation.com/dna/res ... digestion/
http://www.methodbook.net/dna/restrdig.html
by genetherapy
Mon Jul 23, 2012 3:30 pm
Forum: Molecular Biology
Topic: extra unknown band after restriction digestion?
Replies: 11
Views: 9667

Nobody has an idea??
by genetherapy
Wed Jul 18, 2012 5:02 pm
Forum: Molecular Biology
Topic: extra unknown band after restriction digestion?
Replies: 11
Views: 9667

Re: extra unknown band after restriction digestion?

attached other format of the view
by genetherapy
Wed Jul 18, 2012 1:49 pm
Forum: Molecular Biology
Topic: extra unknown band after restriction digestion?
Replies: 11
Views: 9667

Re: extra unknown band after restriction digestion?

<a href="http://tinypic.com?ref=123oztv" target="_blank"><img src="http://i49.tinypic.com/123oztv.jpg" border="0" alt="Image and video hosting by TinyPic"></a>
by genetherapy
Wed Jul 18, 2012 12:18 pm
Forum: Molecular Biology
Topic: extra unknown band after restriction digestion?
Replies: 11
Views: 9667

Re: extra unknown band after restriction digestion?

And yesterday I loaded little amount and looked the gel,did not see any smear but today a loaded whole restriction reaction to gel and checked it and realised have a upward smear. Also have the extra band. Downward smear means degredation but what does upwaard smear. Still thinking about bacterial c...
by genetherapy
Wed Jul 18, 2012 11:48 am
Forum: Molecular Biology
Topic: extra unknown band after restriction digestion?
Replies: 11
Views: 9667

Re: extra unknown band after restriction digestion?

Yes, I checked it. It also have the same extra band
by genetherapy
Tue Jul 17, 2012 6:49 pm
Forum: Molecular Biology
Topic: extra unknown band after restriction digestion?
Replies: 11
Views: 9667

Re: extra unknown band after restriction digestion?

I forgot to say: yes, there was not any smear. Only a band bigger than 10 kb.
by genetherapy
Tue Jul 17, 2012 6:48 pm
Forum: Molecular Biology
Topic: extra unknown band after restriction digestion?
Replies: 11
Views: 9667

Re: extra unknown band after restriction digestion?

I didn't digested e.coli DNA. I purified plasmid dna from e.coli and then digested plasmid DNA. Could it be the e.coli DNA if I could not purify plasmid DNA from bacteria (top 10 e.coli) well. my biggest marker is 10 kb and this extra band is bigger than 10 kb.
by genetherapy
Mon Jul 16, 2012 3:24 pm
Forum: Molecular Biology
Topic: extra unknown band after restriction digestion?
Replies: 11
Views: 9667

extra unknown band after restriction digestion?

I have a vector size 4775 bp and insert 500 bp. I ligated them before and yesterday I digested with related restriction enzymes for my experiment. But interestingly I saw 3 bands. One is insert(500 bp) the other is vector (4775 bp) and there is an other band bigger than 10000bp (My marker's bigger s...
by genetherapy
Mon Jul 16, 2012 3:11 pm
Forum: Molecular Biology
Topic: MOLECULAR CLONING
Replies: 9
Views: 10854

Re: MOLECULAR CLONING

Hi Sameer, The ratio between insert and vector changes whether you have an cohesive (sticky) end insert or a blunt end insert. Generally while we're doing blunt end insert-vector ligation we can make up to 100:1 ratio but while we're doing cohesive end insert-vector ligation generally we prefer 1:1,...
by genetherapy
Wed May 23, 2012 2:49 pm
Forum: Molecular Biology
Topic: ligation problem although try everything,suggestions?
Replies: 7
Views: 7760

Re: ligation problem although try everything,suggestions?

I've bought the ligase 3 days ago. I was using NEB T4 DNA ligase.Now I'm using Quick ligase. I read at somewhere I can't remember now, it says do not spin ligase. Do anyone have experience like this. Can we centrifuge ligase I mean only spin 1 or 2 second? Does it effect it's activity?
by genetherapy
Wed May 16, 2012 3:31 pm
Forum: Molecular Biology
Topic: ligation problem although try everything,suggestions?
Replies: 7
Views: 7760

ligation problem although try everything,suggestions?

Hi all, I'm in trouble with ligation nearly 5 months. My vector is 4775 bp. My insert is 500 bp. I also make PCR to obtain my insert (verify with agorose gel) and then I digested my insert after PCR with the enzymes which my insert have at it's ends (verify again with agarose gel) . Alternatively I ...