im doing a project and i decided to do it on a type of vaccine but i havent made my decidion yet. I wanted to get people’s opininon so thats why im asking this? soooo…. i would really appriciate if u would answer my question. and any doctors out there plz answer. thanks in advance

How did the ingredients of a buffer solution assisted in the extraction of DNA?The experiments purrpose was to extract DNA from a banana. The banana was squished until it was all mushy. Then it was mixed with a buffer solution that contained shampoo,baking soda,salt,and distilled water. I know the shampoo dissolves the lipids and proteins of the cell. During this process the cell membrane is being broken down and the bonds that hold the cell membrane together are filtered out. The baking soda in the buffer solution regulated the pH level of the substance.But can you please help me with the other 2-salt and distilled water. Thanks in advance=)

during DNA purification we do we use alcohol.
would you please explain me the steps in which we can get purified DNA.
+details.
thanks alot
have a nice day
the website says:
Precipitate DNA (salt + alcohol) : For routine precipitation of nucleic acids, you need enough salt to
neutralize the repulsion among the negatively charged strands of DNA, and
enough alcohol to pull water molecules out of the DNA strand.
can you expalin how?
when we lysis enzyme to break down membrane and hen precepitate protien and centrigue them. then we get rid of them.
after that we r left with the crude lysate , and now that it contains the DNA that is (-) charged, the strads will repullse each other. that is why we ADD THE SALT TO REACT with + charges. but then when we extract DNA it will have attached + charged molecules.
+ one more thing: where does the water comes from?? the one that we want to remove using alcohol?
I KNOW IT IS TO MUCH BUT I REALLY NEED TO KNOW WHY : }
THANKS in advance

So I read the glofish website, and it said that
"Do you have to add a fluorescence gene to every fish before it hatches?
No. Today’s GloFish® fluorescent fish are bred from the offspring of fluorescent zebrafish that were originally developed several years ago. Each new GloFish® fluorescent fish inherits its unique color directly from its parents, maintains the color throughout its life, and passes the color along to its offspring."

Then it says
"Does the fluorescence harm the fish?
No. The fish are as healthy as other zebrafish in every way. Scientists originally developed them several years ago by adding a natural fluorescence gene to the fish eggs before they hatched. Today’s GloFish® fluorescent fish are bred from the offspring of these original fish.

Exactly how is the fluorescent protein gene added to the fish?
Every line of GloFish® fluorescent fish (i.e., GloFish® Starfire Red® Zebra, GloFish® Electric Green® Zebra, and GloFish® Sunburst Orange® Zebra) starts with a single fish. The process, illustrated in this chart, begins by adding a fluorescence gene to the fish before it hatches from its egg. Once the gene integrates into the genome (i.e., genetic code) of the embryo, the developing fish will be able to pass the fluorescence gene along to its offspring upon maturity. Because of this, the gene only needs to be added to one embryo; from that point forward, all subsequent fluorescent fish are the result of traditional breeding."

So my question is there any cruelty involved in giving them the gene that makes them glow like there is in dyeing fish? Because I was thinking about getting some, and I wanted to make sure they didn’t suffer at all.

Thanks!! =D
Thanks in Advance!

Can someone explains to me in details about this process and how it works. I tried to do some research on it but i couldn’t find any good source. Thanks in advance.

I’ve been trying to find detailed info on the isolation of plasmid DNA from E.coli cells using phenol chloroform extraction and ethanol precipitation.

If anyone knows of a good website I’d be really grateful if you’d let me know. (the more detail the better!) Many thanks in advance.

When I rack my wine after the primary fermentation, can I put it in a sealed container, or does it have to be in a container with an airlock? Isn’t the only important thing that oxygen doesn’t enter? Thanks in advance from this beginner.

As the title says, tomorrow I plan to have my first attempt at purifyin my his-tagged protein. I finally figured out my ligation issues (DNA was not concentrated enough following the gel extraction). Anyway, I’ve grown up my cells, overexpressed and now they are in the -80C. Tomorrow I plan to break them open and purify. I’ve only run a column on an MBP tagged protein which uses amylose resin in the column.

Is a nickel column for his-tag similar? Is it a resin? If it is, I plan to shake/bind my protein to the resin for an hour before loading it on the column. Do I only elute with imidazole? I know there are plenty of protocols out there but I’m mostly confused about the process. Since this is different than my other purification (3-4 step purification with a few different columns, I’m just not sure what to do). I’ve been told this will be much easier?

I figure step one is to bind my protein to the resin, step two is load it on the column and wash any unbound ptn out. Step 3 is to elute. From there, I can concentrate and also use an optional thrombin cut etc.

OK I drew out this question WAY too long, I apologize. My two questions are:
1) Is it a resin I use for the nickel column?
2) What steps do I need to save for a gel? (Crude, wash, and eluted protein)? Maybe more if I cut the tag?

Thanks in advance

I’ll be transfecting human dermal fibroblasts with a gene in the pIRES2-DsRed-Express vector. I’ve never used a fluor. microscope before and although I know the wavelengths, all the filters are simply labeled by color. Do I use the "red" filter and will the cells containing the fluorescent protein appear red under this filter? Any links that have basic information on these types of procedures will be greatly appreciated.
Thanks in advance.

(I’m just sick of looking up information, can’t find anything relevant.)
How can scientists turn genes on and off? Also, what are some implications of this? In other words, if scientists figure out a way to control expression of an introduced gene, how might this affect the medical community and the field of gene therapy?

Thanks in advance!