Search found 18 matches

by citroenboom
Wed Nov 14, 2012 1:44 pm
Forum: Molecular Biology
Topic: molecular cloning failure
Replies: 3
Views: 3232

Re: molecular cloning failure

Some more data would be helpfull.
What kit did you use to get the DNA? How much did you load on the gel? Did you see the marker? Did you see other bands in the gel?
by citroenboom
Wed Nov 14, 2012 1:41 pm
Forum: Molecular Biology
Topic: separation problem with GelRed
Replies: 12
Views: 34613

Re: separation problem with GelRed

Sadly I did not help us out. In our lab often we want to check early if we have good product/restriction pattern. And with GelRed gels have to run longer to get nice bands.

But it helps indeed if you run longer. Then result can be nice.
by citroenboom
Wed Nov 14, 2012 1:37 pm
Forum: Molecular Biology
Topic: Sequencing question/primer design
Replies: 4
Views: 5520

Re: Sequencing question/primer design

Do not focus too much on Tm = 58. A primer of about 20 to 25 bases should do the trick. I always make sure there is a C or G at the end. Longer primers naturally give more trouble like hairpins. For checking my primers I like to use:
http://basic.northwestern.edu/biotools/oligocalc.html
by citroenboom
Wed Nov 14, 2012 1:32 pm
Forum: Molecular Biology
Topic: E.coli Genotype
Replies: 4
Views: 6205

Re: E.coli Genotype

According to openwetware it blocks metabolism.
I am using TOP10 and it has the same deletion. My cells do happily grow on M9 or the minimal medium of our group with leucine. DH5a doesn't have it I think.
by citroenboom
Wed Nov 14, 2012 10:50 am
Forum: Molecular Biology
Topic: Cloning software
Replies: 1
Views: 2681

Cloning software

For a practical course we are searching for a program that we can use for cloning in silico. Like Clonemanager or VectorNTI. We cannot afford to buy the programs for the course and we would like to show the students how the cloning works. They will do the task while the transformation runs to reduce...
by citroenboom
Mon Sep 03, 2012 11:45 am
Forum: Cell Biology
Topic: Why do cells age?
Replies: 22
Views: 23669

Still not all cells in a colony will grow. If I remember correctly from a presentation here some cells stop growing after a while and die. Or turn to 'stand-by modus.

A complete colony contains so much cells that some can survive, but most will not. It is a matter of selection.
by citroenboom
Mon Sep 03, 2012 11:40 am
Forum: Molecular Biology
Topic: How to detect proteins in my sample?
Replies: 3
Views: 3289

Just look for SDS-PAGE. And you can combine both techniques :)
by citroenboom
Mon Sep 03, 2012 11:36 am
Forum: Molecular Biology
Topic: Vector
Replies: 2
Views: 2924

Re: Vector

Maybe check the marker? Then you will find the amount of basepairs.
by citroenboom
Tue Jul 17, 2012 9:31 am
Forum: Molecular Biology
Topic: restriction analysis
Replies: 14
Views: 14723

If your PCR is successful (check 5 ul on 0.8% agarose after the run) you can load the whole PCR mix on gel and cut it out. I prefer to use PCR-purification kit, but both works. So, yes, I think your procedure should work fine.
by citroenboom
Tue Jul 17, 2012 7:29 am
Forum: Cell Biology
Topic: Why do cells age?
Replies: 22
Views: 23669

And what about my precious E. coli! They do not contain telomers :)

They age because of damage by the environment, by dividing, DNA damage and that sort of things.

In Groningen University (The Netherlands) is some nice research going on about this topic (as I understood)
by citroenboom
Tue Jul 17, 2012 7:26 am
Forum: Genetics
Topic: Explain why Genetic Code is called "Rosetta Stone" of life?
Replies: 11
Views: 17964

You seem to understand the principles of the genetic code. But, sorry, you failed coupling it to the Rosetta stone. Hope you passed the test. This stone appeared to be the key to understand several languages. Just as the genetic code is the key to understand the way life functions and the aa sequenc...
by citroenboom
Tue Jul 17, 2012 7:10 am
Forum: Molecular Biology
Topic: restriction analysis
Replies: 14
Views: 14723

For PCR you need a template = your gene of interest. For this you can use your vector with your gene in it. You design your primers to match the beginning and the end of your gene. Also add the sequence of the restriction sites you want to use, and add GATCTAG to the end of both sides (to give the r...