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Totipotent Stem Cell From Molecular Components

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Totipotent Stem Cell From Molecular Components

Postby caters » Sun Feb 02, 2014 10:58 pm

Q1 I am wanting to make an animal cell from DNA, RNA, Protein, and Fatty Acids as well as Glucose and other sugars. I am wondering. Should I start with the cell membrane or the nucleus and nucleolus(both are very important)?

Q2 The pace of DNA replication is 8 2/3 hours for every chromosome and 2097152 of each chromosome in 1 week. Is that just an estimate because I know that it is 8 2/3 hours per replication origin.

Q3 Once I know how many proteins of a particular type should I do this:

Go to BioGPS

Look up the proteins

Make sure I have human selected(since I am making a human stem cell)

Average the mRNA expression to get stem cell mRNA expression

Multiply by 100 to get the amount of protein

Q4 I have heard of amplicons used in PCR. Do I really need one to copy a whole gene or just primers, DNA, and enzymes?
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Re: Totipotent Stem Cell From Molecular Components

Postby JackBean » Tue May 13, 2014 5:04 pm

A1: good luck, even Venter haven't managed that yet.

A3: and how will you get the protein? Are you really sure that the stemm cells contain all protein in the same amount?

A4: amplicon is the result of PCR. You need some template though, in your case probably the DNA.
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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Re: Totipotent Stem Cell From Molecular Components

Postby caters » Fri Aug 01, 2014 1:42 am

I am not saying # of protein x = # of protein y(In fact this is almost never the case). Rather I am saying that for a particular protein in my stem cell at least, the amount of that particular protein is the average of the gene expression in # of strands of mRNA * 100.

I am getting the protein this way.

I take the results of PCR and put it in a test tube with RNA polymerases, Ribosomes, and other proteins that help with protein synthesis. I then look at the structure of the protein as is and inside a cell(every single bond) and change the structure if needed. Last but not least, before I put the protein in the organelle I separate it from the protein synthesis enzymes using a centrifuge.

As far as the DNA side of things first I do DNA extraction(in this case of my own DNA). I then separate the chromosomes using differential centrifugation. Next I put it in an incubator with the same DNA replication enzymes as the ones in the cell at human body conditions and leave it there for 1 week where it replicates at a pace of once every 8 2/3 hours. I then separate a diploid set from the rest of the DNA and use the rest of the DNA for PCR. I form the histone proteins at the same time that I am forming the nucleus and put the histone proteins in the test tube with the diploid set of chromosomes, make sure there is no more than 1 chromosome per histone, and use telomerase to extend the telomeres before I put the diploid set of DNA in the nucleus.
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