Greetings, here is friday morning!
Nothing but, there is a question as to the microarray. I use Arrayit micro spotting solution 2X plus. As you can see in the photograph, after baking recommended in the protocol, we saw a dried donut shape. And the fluorescence also appeared as a donut shape. It seems to have dried coffee ring effect in dry environment.
I am using the dry oven because I do not have enough money to equip it, but I have to keep the humidity to some extent by looking at the protocol of the arrayit. Is there any way to keep the humidity in the dry oven?
When I kept the humidity at 100%, I could not see any fluorescence(when I used 50% DMSO). Is there any other way to prevent the effect of coffee ring and spotting it in a circle?
PS: Fluorescence is very weak. Should I increase the concentration of the target DNA in this case, or should I increase the concentration of the probe in proportion to that?
I used a small airtight box of plastic to hold the humidity.
Something like this hyb chamber in the picture. You can also use a 50ml falcon.
And you should place some water within the chamber.
The dots on my first slides looked similar. I have to look in my old records.
@Björn Abendroth Nope, I baked right direction(spotted->up). I spot 1ul DNA(0.1ug).
Ok.
What kind of slides do you use?
Are there activ groups on the surface were you bind the probes?
@Björn Abendroth Arrayit superamine2.. (oligo solution is arrayit micro spotting solution 2X plus) it coated by aminosilane.. Oligonucleotides aren't modified because it can immobilize DNA by UV or baking.
Ok. I have look in my records. Maybe i will find something.
I have seen the article you posted "Hybridization-induced suppression of coffeesharse effect for nucleic acid detection".
But this is not the methode you working on?
Just to be sure.
We are talking about a normal microarray with spotted probes ,to which the labelled DNA template should hybridize?
Regards
Björn
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