What does hot water do to the cell during an DNA extraction?
The main steps of DNA extraction are cell lysis (where the cell wall and membrane are ripped apart), seperation of DNA from the rest of the cell contents (proteins, RNA) and the final precipitation and wash of the DNA for purity. I am not sure what protocol you are talking about, but if you are boiling the prep, you are probably lysing the cells. This is also, and usually achieved through the use of detergents and lysis enzymes (lysozome or proteinase K).
Can any one explain this DNA extraction protocol i have few questions?
1) your RNA is in along with your DNA. This protocol does NOT separate DNA from RNA. For some downstream applications that doesn't matter, but for others, you may want to add RNase to your lysate about 30-60 min before you precipitate the proteins--that'll get rid of the RNA. 2) After your final 70% ethanol wash, your DNA will be an insoluble pellet at the bottom of your tube. You can't do anything with that DNA unless it's in solution, so you add water or buffer (typically Tris-EDTA [TE], pH 7.4-8.0) to hydrate and solubilize your DNA. TE buffer is better than water, since DNA is more stable in TE than in water, which has a lower pH. 3) 100% ethanol removes ALL the water molecules, which might make the DNA harder to re-solubilize, so theoretically, the 70% ethanol wash will make it easier to resolubilize the DNA. However, in my own experience, 100% ethanol CAN be used, and the DNA will STILL resolubilize in TE buffer--It just takes a little longer than if I use 70%.
Do you think the same protocol for DNA extraction would work on bacteria? Please explain..?
It's been a long time since I extracted DNA lemme think... I would have to guess 'NO' because wheat germ is from a plant and therefore eukaryotic, while bacteria are prokaryotic. So you might need or want to separate out nuclei in the wheat germ extraction where these would not exist in the bacterial extraction. Plant cells have a cell wall, made of cellulose, which would have to be dissolved in some way to extract the DNA. Bacteria may have various substances surrounding them (or no hard wall as well). There's other differences. The protocol would be vaguely the same. There's a subset of the steps that would be the same. But all the steps the same? Doubt it, unless the bacteria are very plantlike.
How long does DNA extraction take?
I registered a DNA kit through 23AndMe and it arrived on Dec. 25th, 2017, And they’re still on the DNA extraction stage. How long does it normally take? I was informed that the whole analysis could take up to 8 weeks, but I’m just wondering what they do and how long it takes to extract and purify the DNA from your saliva.
What's a dumbed-down definition of DNA? Like - 8th grade dumb? ?
Well there is no dumbed down definition of DNA: DNA is a short form for dioxyribonucleic acid, it is a nucleic acid that contains the genetic instructions used in the development and functioning of all known living organisms and some viruses. In your dumbed down version, we can say that DNA contains the genetic information that codes for characteristics for humans. We can also see that DNA is composed of subunits called nucleotides: A nucleotide is the repeating units of DNA which consists a phosphate group, sugar (deoxyribose) and a nitrogenous base. Again, the dumbed down version can be understood by saying in DNA, there is a repeating structure of nucleotides, which contains the coding for the genetic information.
My phenol:chloroform:isoamylalcoh... is pink. Is it normal? I just can't seems to extract any DNA...?
Yes, your p:c:i can be pink. This is not unusual. There could be many things that are going wrong. Are you trying to purify total DNA or plasmids? How are you lysing the cells. I can't tell based on your protocol. You don't specify how you are lysing the cells. I am not sure what STE buffer is so my apologies if this is how you are lysing your cells. Assuming you lysed your cells, removed all cellular debris and resuspended your DNA/RNA in TE, make sure that your TE isn't acidic. If it is, your DNA is sitting in the organic phase. Under acidic conditions, DNA will stay in the organic phase while RNA will be in the aqueous phase. This is a common way to extract RNA from cells. Based on your protocol, this would seem to be the most obvious thing going wrong. Where did you get this protocol? From a reference or homemade? If I could look at the reference and get a little more detail, I might be able to help more.