Search found 9 matches

by chaka8
Thu Jul 31, 2008 10:26 pm
Forum: Molecular Biology
Topic: Morpholino review: how to knockdown
Replies: 7
Views: 18038

Re: Morpholino review: how to knockdown

Jon, Thank you for your detailed and illuminating response. As I said in my first post, MO is an excellent tool, in many cases the only tool available. As a researched working on ZF CNS development, I see many MOs non-specific effects that make research very difficult indeed. For this reason I feel ...
by chaka8
Thu Jul 31, 2008 3:43 am
Forum: Molecular Biology
Topic: Porblems making DIG probe - please help
Replies: 5
Views: 7061

Re: Porblems making DIG probe - please help

OK, did the whole thing over again, everything new. I ran a non-denaturing ethidium bromide gel. In one lane I ran just some of the DIG RNA mix and I see nothing, shouldn’t I see a strong band of the free RNA neucleotides?? In the other lanes I ran 1ul of the reactions (after 2.5 hours at 37c), I do...
by chaka8
Wed Jul 30, 2008 4:12 pm
Forum: Evolution
Topic: Age of Man
Replies: 31
Views: 19329

Re: Age of Man

Going back to the original topic, fissile dating is by no means the only way to estimate the immergence of humans. Genetic calculations based on rate of mutation is also an excellent method. Time frame of divergence between apes and modern humans from a common ancestor can has also been calculated b...
by chaka8
Wed Jul 30, 2008 12:52 pm
Forum: Evolution
Topic: Offspring Selfishness changing into Parental altruism?
Replies: 5
Views: 6495

Re: Offspring Selfishness changing into Parental altruism?

However, If I was to generalize I would say that in infants, that are not fully developed in terms of hormones, brain structure, cognition etc, there is no grasp of altruism. Evolutionarily speaking it makes sense that infants, that are competing with siblings and have no real use for altruism woul...
by chaka8
Wed Jul 30, 2008 12:39 pm
Forum: Molecular Biology
Topic: Morpholino review: how to knockdown
Replies: 7
Views: 18038

Re: Morpholino review: how to knockdown

Snowcapk, As far as I am concerned the gold standard is rescue. One way is to make mRNA using the Mmassage Mmachine kit and inject with your MO, if you are lucky it will rescue the phenotype and this means that the MO has a specific effect on the target gene. However, more often than not this will n...
by chaka8
Tue Jul 29, 2008 4:47 pm
Forum: Evolution
Topic: Offspring Selfishness changing into Parental altruism?
Replies: 5
Views: 6495

Re: Offspring Selfishness changing into Parental altruism?

These are many genes that change and even reverse their function with the age of the individual. An example is GABA which switches from being inhibitory to being excitatory with age. Regarding altruism, I think that you are oversimplifying. Altruism is a complex behavior that is no doubt modulated b...
by chaka8
Tue Jul 29, 2008 1:30 pm
Forum: Molecular Biology
Topic: Morpholino review: how to knockdown
Replies: 7
Views: 18038

Re: Morpholino review: how to knockdown

With all due respect, I don't think that an article about morpholino written by people that work for the company that makes morpholino would be a good place to start anything. I'm actually surprised that this was allowed. “specific, and lack non-antisense effects"? They must be joking! I use mo...
by chaka8
Tue Jul 29, 2008 11:42 am
Forum: Molecular Biology
Topic: Porblems making DIG probe - please help
Replies: 5
Views: 7061

Re: Porblems making DIG probe - please help

Thanks snowcapk, Haven’t tried the control, I don't think that the Roche kit comes with one but I'll check. I'm sure about matching the polymerase to the liberalized vector. The DIG mix contains everything and it’s the RNA mix. I know that polymerase buffers have a precipitant and I vortex and even ...
by chaka8
Tue Jul 29, 2008 2:02 am
Forum: Molecular Biology
Topic: Porblems making DIG probe - please help
Replies: 5
Views: 7061

Porblems making DIG probe - please help

Hi, I am having problems with making DIG RNA probes (Roche). I get no bands when running a gel after purification other than the weak band of the linearised plasmid. I am using T7 and SP6 polymerase. I seem to have narrowed down the problem to the process of making the probe (I have no band before p...