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On another newsgroup, http://groups.msn.com/aidsmythexposed/general.msnw, there was a debate about whether "Hepatitis C" virus had been isolated. I will quote a few passages below, and if anyone else wants to get involved, feel free, provided that you arguments are on point and supported by evidence. The posts below are from the "middle" of the debate, which involved others (not just the two quoted below, "Softrat" and "Biolad"):
Softrat:
QUOTE: Here they are:
http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&uid=11314272&cmd=showdetailview&indexed=google
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=334354
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1370611
http://linkinghub.elsevier.com/retrieve/pii/S0014579302028120
http://www.pubmedcentral.nih.gov/articlerender.fcgi?&pubmedid=12584327
Look! What Dr. Lanka was talking about!
"When RT switches templates during cDNA synthesis, it creates a progeny cDNA sequence that harbors sequence identity to both templates"
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WG1-4J5T5V7-1&_user=10&_coverDate=07%2F31%2F2006&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=5792be01e26c83d47188af2c0f4b8d17
This stuff is all over the net! Just use your search engines.
Bio Lad said:
"First, a cDNA is a DNA copy generated from an RNA template. You must therefore have an RNA template to begin with. To claim they generated cDNA without having isolated RNA is just silly. It is like saying you made a photocopy of a page without using the original paper"
Yes many little mRNAs you have isolated and then template switched.
"They then reverse transcribed the RNA and THAT is how they made their cDNA. The cDNAs they obtained are nothing more than DNA copies of the RNAs they isolated. "
But cDNA is made from template switching. Like I said.
"Nope, once again it is your misunderstandings and assumptions that lead you to believe that. Especially since the paper describes the RNA isolation (M&M, paragraphs 1 and 2)."
Once again cDNA is made from template switching. UNQUOTE.
And:
QUOTE: This is fromthe study Biolad cited:
"A lambda gt11 cDNA library was constructed from RNA purified from hepatitis B viral surface antigen-negative human plasma with high alanine aminotransferase activity. A cDNA clone, designated as C8-2, was isolated by immunoscreening with mixed sera from non-A, non-B hepatitis (NANBH) carrier and convalescent chimpanzees"
Now look at the quotes in the above post. Pay note to this one:
"Here, we describe a fast, simple method for constructing full-length cDNA libraries using SMART technology. This novel procedure uses the template-switching activity of Moloney murine leukemia virus (MMLV) reverse transcriptase to synthesize and anchor first-strand cDNA in one step."
It looks like template switching is how cDNA is made. UNQUOTE.
Biolad:
QUOTE: SoftRat,
Your reasoning on the issue of template switching is not as relevant as you would think. I am in the process of typing up a full rebuttal using, amongst others, the citations you provided. Since, however, you seem to have a history so far in this debate of making assumptions based on misunderstandings, please take this time to look back through your sources and try to see if you can figure out why your argument it not valid. I should have my post sent tonight as I am busy at work today. Keep in mind that my posts all go through Rod first so it may take a while to actually get posted so they generally have a bit of a lag between the time I send it and the time it gets posted. It is understandable though as I'm sure he has other things to do as well.
-BioLad UNQUOTE.
Softrat:
QUOTE: What we have learned Part 2
<o:p></o:p>
By: The Soft Rat
<o:p></o:p>
Ok very interesting stuff.
<o:p></o:p>
Bio tried to argue that I was wrong about cDNA being made via the template switching method. But it turns out I was in fact completely right about this. He also claimed that Dr. Stephan Lanka, some with a PhD in molecular biology and who has isolated his own phage virus old school style, was wrong about viral cDNA being made artificially through reverse transcriptase template switching.
<o:p></o:p>
But as these quotes show Bio Lad appears to have been misleading us. Remember Bio Lad has a masters in molecular biology so he is well aware that cDNA is made via reverse transcriptase template switching. Which means that him trying to mislead us this way is no honest error on his part.
---------
The Internet source for these quotes are listed in above post and it is easy to find this stuff on the net with a search engine:
<o:p></o:p>
“When RT switches templates during cDNA synthesis, it creates a progeny cDNA sequence that harbors sequence identity to both templates"<o:p></o:p>
<o:p></o:p>
Real important one here as this is not only what Dr. Lanka was talking directly about but it shows that cDNA is made from many small pieces of mRNA.<o:p></o:p>
<o:p></o:p>
“cDNA strands produced from template switching were selectively amplified�?lt;o:p></o:p>
<o:p></o:p>
“Temperature-dependent template switching during in vitro cDNA synthesis by the AMV-reverse transcriptase....�?lt;o:p></o:p>
<o:p></o:p>
More proof that cDNA is made via the template switching method.<o:p></o:p>
<o:p></o:p>
This next one is important as it shows that template switching creates fake genomes:<o:p></o:p>
<o:p></o:p>
“Evidence that BmTXKβ–BmKCT cDNA from Chinese scorpion Buthus martensii Karsch is an artifact generated in the reverse transcription process. "<o:p></o:p>
<o:p></o:p>
<o:p></o:p>
“Because it is known that reverse transcription (RT) can produce artefacts by strand switching during the RT step�?�?BR>
<o:p></o:p>
This last quote is from the scorpion paper at:
<o:p></o:p>
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1370611
<o:p></o:p>
“Here, we have investigated the presence of such template-switching artifacts in
cDNA databases�?lt;o:p></o:p>
<o:p></o:p>
<o:p></o:p>
So cDNA is made via template switching that does create cDNA genomes that do not correspond to real world genomes. It creates fake genomes.
<o:p></o:p>
Bio Lad didn’t tell us any of this. He clearly did everything he could to steer us away from template switching and deny that the Hep C cDNA was made via template switching. This is no over site on the part of Bio Lab. He has a Masters in molecular Biology. He knows exactly what he is doing here.
<o:p></o:p>
The critical and fundamental part of my argument is that the cDNA of Hep C is made via template switching from smaller unrelated mRNA molecules. And therefore the Hep C mRNA is not found in nature but is a Lab artifact.
<o:p></o:p>
Take the time to read through the study Bio Lad sited. There is not one shred of evidence that they isolated a single molecule of mRNA and somehow made the cDNA from that. Instead if you read the materials and methods section you will see the just extract out mRNA from the plasma. There is no proof the Hep C mRNA is in there as a single entity. From there they make cDNA via reverse transcription. And as the evidence above shows cDNA is made via template switching with reverse transcription enzymes. Just like I was saying.
<o:p></o:p>
http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=330752&blobtype=pdf
<o:p></o:p>
<o:p></o:p>
Now Bio Lad said:
<o:p></o:p>
“They then reverse transcribed the RNA and THAT is how they made their cDNA. The cDNAs they obtained are nothing more than DNA copies of the RNAs they isolated�?lt;o:p></o:p>
<o:p></o:p>
Notice how tricky he is. He tells lies of omission. He doesn’t deny the template switching directly, he just fails to mention it. However given the fact that he is rebutting my template switching argument he is clearly imply that template switching is not happening. But cDNA is made by template switching.<o:p></o:p>
<o:p></o:p>
“Nope, once again it is your misunderstandings and assumptions that lead you to believe that. Especially since the paper describes the RNA isolation (M&M, paragraphs 1 and 2)."<o:p></o:p>
<o:p></o:p>
Once again he fails to mention that they extract or isolate mRNA, but there is no proof that the mRNA they extract contains any mRNA existing in the form of the Hep C virus. And given the fact that they then make cDNA which is made via template switching it is clear that the sample must not have. The extracted sample must have been composed of many small pieces of mRNA that were then template switched together into the cDNA.<o:p></o:p>
<o:p></o:p>
Now if Hep C virus really does exist why is Bio Lad, someone who has a masters in molecular biology, misrepresenting the way cDNA is made? Why is doing everything in his power to steer us as far away as possible from understanding the fact that cDNA is made via template switching with reverse transcriptase enzymes?<o:p></o:p>
<o:p></o:p>
Remember my argument that Hep C mRNA does not exist, and thus Hep C virus does not exist is based on the fact that cDNA is made via template switching. The very thing Bio Lad tried to mislead us into believe was not true. I think the facts speak for themselves.<o:p></o:p>
<o:p></o:p>
Also do not forget that Bio Lad failed to inform us that when the human, or chimp, DNA genome database is made they automatically remove any and all evidence of viral genomes in there. So this is why when you do a BLAST nothing shows up. Once gain why did Bio Lad fail to mention this to us?<o:p></o:p>
<o:p></o:p>
I think Bio Lad is someone that came here to stop all this talk about viruses not existing. I don’t know if he came here on his own, or if some company, or the NIH sent him or what. But it is clear that this is what he is trying to do. It is also clear that he will distort the evidence as needed to do this.<o:p></o:p>
<o:p></o:p>
This is not consistent with Hep C or viruses in general exist. If they did exist Bio Lad could reveal all relevant data on the subject matter and let reality speak for itself.
------
Update:
Bio lad said this:
"Your reasoning on the issue of template switching is not as relevant as you would think."
If this is true Bio Lad then why were you not up front about the fact that cDNA is made via template switching? Why did you clearly imply that I was off base on this?
Bio lad you are hiding relevant information here. As a molecular biologist you should not have to do that if Hep C virus really does exist. UNQOTE.
And:
QUOTE: Bio Lad:
I told Rod Knoll I would stick with you till Monday. When I get home tonight it will be Tuesday Morning. So this will be my last post on this Hep C series.
You are now about to construct an argument to show that my reasoning on template switching is not as relevant as I think it is.
However you felt no need to construct any such argument until I revealed that you had been hiding the fact that cDNA is in fact made via template switching via reverse transcription enzymes. This clearly shows that you where covering up that the Hep C genome is created from smaller mRNA sequences strung together via template switching. Now that I have showed this is the case you will cinstruct an argument that shows this is not really as relevent as I think it is.
Bio Lad in order for you to be regarded as a valid expert on molecular biology, and the existance or non-existance of Hep, we have to be able to trust what you say. We can't regard you as a reliable witness if we have to worry about if you are telling us everything we need to know in order to come to a relaity based conclusion.
As an expert in molecular biology, and a researcher, you have hidden two very imporatnt things that you would clearly have known about. One is that I was dead on the mark about cDNA being made via template switching. The other that the reason Hep C sequences can not be found in the data base when you BLAST is because they would have been removed, or never added, from the data base when they were found.
Because you have hidden these things is does not matter what you say in your next rebuttle. You simply can not be trusted. If I looked through your rebuttle and worked long and hard on it I might be able to uncover how you are fibing or hiding something important and then I would post that here. The only problem is you would then construct a new argument showing how this new information I have uncovered, and that you were hiding, is not realy as relvant as I think it is. I do not care to find out how long you would be willing to carry this on.
I have had to put aside other things I would rather be working on in order to have this debate. The fact that you have been hiding the fact that cDNA is made via template switching means you have been wasting my time. The fact that you do not reveal relevant facts, and pretend they do not exist, and only then deal with them after I show you have been hiding them means your testimony about Hep C carries a value of zero epistemologically. If I was on the jurry in court I would have to disregard your testimony as there is no way for me to know when you are telling the truth or not. No way for me to know if you are telling me everything I need to know to come to a valid conclusion.
Also lets look at this:
"First, a cDNA is a DNA copy generated from an RNA template. You must therefore have an RNA template to begin with. To claim they generated cDNA without having isolated RNA is just silly. It is like saying you made a photocopy of a page without using the original paper. Second, The Chiron group did in fact isolate the RNA from serum. They state it quite clearly in the first page under materials and methods (second paragraph). After obtaining the RNA they state:
"The RNA was further purified by DNase treatment (1. 15KU/mlDNase, 50 mM Tris-HCl pH7.4, 1 mM EDTA, 10 mM MgCl2)
for 30 min at 37°C, and anion exchange chromatography (Qiagen pack-100, Diagen)."
Since you seem sketchy on your molecular biology, and for the benefit of anyone not familiar with it, DNAse is an enzyme that degrades DNA into single base units. This means that any DNA that may have been present in the serum is now completely degraded and no longer usable as a template. They then reverse transcribed the RNA and THAT is how they made their cDNA. The cDNAs they obtained are nothing more than DNA copies of the RNAs they isolated. (btw, in case you are about to make another bad assumption, the DNAse is heat inactivated prior to reverse transcription.)"
Notice you never directly say that the cDNA is not made via template switching. You just fail to mention it. You fail to mention it in this post or any others even though you know I have been saying the cDNA is made via template switching. You fail to mention this fact in any post even though you know the template switching is the fundamental essense of my argument that the Hep C genome is made from smaller mRNAs strung together in template switching, and thus the Hep C mRNA genome does not exist in nature.
You only now want to deal with this fact after I have shown that cDNA is made via template switching. Befoe this you were more tha happey to keep this under wraps. You are not a trustworthy source of informationon this isuue Bio Lad. There is no way anyone can know when they read yournext post if you are being honest with them or not. Thus your testimony must be thrown out of court.
I do not know what else you may have hidden from us that I or others have not yet uncovered. But let me tell you this Bio Lad. I have dusted off my old college BioChem text book. Since this was written some time ago I will be going to the UW and also buying a new text on molecular biology so I can have up to dat information. So given a few years I will catch up to you.
Also like I told you I am studying virology from begining to end. Oneof the reasons I am doing this is cause the early research on virology is on my level. As I move through the papers I learn the methods of molecular biology as the scientist of those days developed them. So I learn as they learned. In the order that the methods were developed.
I also get to see how the belief in replicating particles started. I get to read the papers from those days that reported evidence against virology. I and guess what Bio Lad? In those days there were scientist that disagreed with the beleif in viruses. There where scientist that showed that the disease causing agents did not self replictae. That the sap of plant viruses did loose potency as it was transfered from pant to plant. That high doses of that sap produced strong effects, and low dosses produced weak effects, just like a finite toxin would. I am working on a paper write now that goes into all of this and documents all the research papers on this matter. So given a couple years I will get back to Hep C. UNQUOTE.
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