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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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Postby chance » Wed Sep 09, 2015 5:35 am

PCR is an in vitro DNA amplification technology, which is dependent on the DNA polymerase chain reaction, which is the three step reaction of DNA polymerase chain reaction, which makes the DNA fragment increase in number and the number of specific gene fragments in short time.
The basic principle of PCR is similar to the natural replication process of DNA.
http://www.biologicscorp.com/
http://www.biologicscorp.com/
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