RNA:DNA Hybrids Survive Digestion in mRNA Vaccine Manufacturing
New paper LIVE today that proves that the production process of modRNA was highly flawed in addition to HOW and WHY the DNA is in the vials and was not detected/reported
First things first.
The paper is entitled: “RNA:DNA Hybrids Survive Digestion in mRNA Vaccine Manufacturing”. It was published on January 13, 2026 in the Journal of Independent Medicine1 and it is about DNA being in the nucleoside modified RNA-LNP COVID shot vials (Pfizer/BioNTech and Moderna) - in copious amounts - and it details why it’s in these vials. It verifies our hypothesis that the DNA in the vials escaped enzyme degradation during the production process because the wrong enzyme was used to clean out the DNA at the end of the process. This wrong enzyme called DNase1 could never degrade RNA:DNA hybrids - which inevitably form during the in vitro transcription (IVT) process - and the manufacturers knew this.
Here’s a reminder of the modRNA synthesis process used as part of Process 2.
The DNA template is the exact complement of the RNA produced (newly synthesized) during the IVT process, so naturally, the RNA strand can politely ask the [non-template strand] from the double-stranded DNA to scooch over to replace it. When the RNA and DNA strands stick together, which they inevitably do, they stick together hard. This is because the N1-methylpseudouridines have a high melting temperature (Tm) and thus require high temperatures to rip apart. They be sticky.2
And, by the way, when I say the manufacturers knew this, I mean it.
“The specific activity of DNase I for RNA:DNA hybrids, however, is at least 100-fold below that for dsDNA (Sutton et al., 1997).”34
BioNTech SE, Mainz, Germany
Wait, what? A BioNTech team published this in July 2024?
In fact, all of the authors on this paper worked for BioNTech.
The authors declare that this study received funding from BioNTech SE. The funder had the following involvement in the study: all authors are current employees at BioNTech SE.
I want to remind everyone that this is yet another point during the manufacturing/production process where the manufacturers did more than a big oopsie. Substituting in N1-methylpseudouridines into every single U position (there are 801)5 was a big mistake that lead to many problems such as frameshifting, and as is suggested in our paper, codon optimization for deposition of coding material into humans via LNPs leads to the significant enrichment of GC content.678
The reason that it is so significant that they used DNase1 instead of say, DNase-XT for degradation of potential RNA:DNA hybrids, is two-fold:
They knew DNase1 wouldn’t degrade potential hybrids efficiently and that any hybrids would inevitably be packaged into the LNPs, and
This would lead to inevitable delivery of these cancer-inducing foreign molecules into the cytosols, and perhaps even the nuclei, of cells.
Did they not know the impact of introducing foreign RNA:DNA hybrids to human cells by transfection? Did they not fully understand that R-loops directly cause disease in humans when accumulations occur?
R-loops do occur naturally in our cells (mostly during transcription) and can serve physiological roles like aiding gene regulation or replication, but they can also be pathological when persistent (or accumulative) leading to DNA damage (cancer), replication stress, or inflammation.9 So can you imagine - on top of normal ongoing physiological processes - what your poor cells have to deal with when introduced to hordes of these additional foreign molecules. Wouldn’t anyone with half of a brain cell anticipate R-loop accumulation that might lead to disease states like cancer?
The other can of worms that our paper clearly describes is how the manufacturers had tests for Spike DNA detection10, but only used tests to find KAN DNA. It’s like “hide the Spike game” - en masse. Except this isn’t a game, is it?
A little about the KAN gene
The KAN antibiotic resistance gene is the gene encoding neomycin phosphotransferase II (never mind) which confers resistance to [the aminoglycoside] antibiotics kanamycin and is often used as a selectable marker in molecular biology plasmids to identify successfully transformed cells → like our infamous E. coli colonies grown on kanamycin-containing media during the N1-modRNA-IVT-synthesis/manufacturing process.
As you can see from Figure 1 in our paper, those DNase1 enzymes that they used at the end of the IVT synthesis process did not work on RNA:DNA hybrids. They did, however, work on the other DNA elements from the plasmid that were not designed to produce modRNA - like KAN DNA.
Do you see the problem?
Here’s a quiz for you guys: If you were an evil genius and you wanted to hide high potentially high DNA levels from regulators, what would you do, and what kind of tests would you run, to show low DNA levels? Maybe you even want to show high RNA levels.
I’ll give you a big hint: According to a Therapeutic Goods Administration (TGA) document (Residual DNA Quantitation in Moderna mRNA Vaccines by qPCR), Moderna designed and used a qPCR detection method for KAN DNA.11 They have one for Spike as well, so why didn’t they use it?12 My guess would be because they knew the KAN DNA results would come up low to nil and that way, they could claim that there’s no DNA in the vials. Presto magico!
The Moderna SOP-1020 can be found here, and notice that it was written on October 9, 2020.
This document describes the method used for quantitating residual DNA in Spikevax mRNA vaccines by qPCR.
Residual DNA, eh? Well, not ALL residual DNA, right guys?
Why would Moderna explicitly outline a qPCR assay designed to amplify the KAN gene in 2020? And furthermore, isn’t it weird that this would only be useful in the context of a plasmid/E. coli IVT synthesis pathway like the one they eventually upscaled to - aka: Process 2? Are they psychic? Why was this assay developed during Process 1 use, as in, during the Pfizer phase III clinical trials?
Perhaps to show the eventual lack of contaminating DNA presence? Their DNase1 enzyme worked fine on KAN, so, no problem, right?
Wrong.
If they had used (and I suspect that they did) a qPCR Spike assay to check for contaminating Spike DNA, they would have (and likely did) find loads of it. Even above their precious 10 ng/dose EMA level.
So what did our paper report overall? Loads of DNA, that’s what. And we were able to verify why we found it: it’s because of the hydridization of “vaccine” RNA to DNA templates during production for which their pre-determined enzymes could never work, and for which their pre-determined “tests” would never measure.
Our findings reveal that residual DNA quantification in mRNA vaccines varies by more than two orders of magnitude depending on the choice of assay target and nuclease treatment conditions. This discrepancy is not merely a technical artifact but reflects a fundamental biochemical reality: RNA:DNA hybrids formed between vaccine mRNA and template DNA resist standard DNase I digestion. When regulatory assays rely exclusively on amplifying DNase-sensitive loci, they systematically underestimate the total burden of residual plasmid DNA.
So they made a KAN assay back in 2020 to pick up plasmid-derived KAN and they knew it wouldn’t pick up any significant amount of contaminating DNA because the DNase1 enzyme would have chewed it up sufficiently. OK.
Let’s look at Figure 3 in our paper which shows exactly how and why we know we’re right about the RNA:DNA hybrid theory.
In the figure below, you can see are qPCR readouts for Ori and Spike DNA from the Moderna and Pfizer vials we tested. As a reminder, qPCR means quantitative PCR and it is used to detect DNA fragments (or to produce amplicons) using specifically-designed primers. These primers “capture” and amplify specific fragments of DNA that one might suspect is in a given sample. The specificity comes entirely from the primer design and ensures that only the suspected/target fragments are amplified and detected. By the way, this is why using fluorometry (in tandem) to measure all DNA is so important.
The green lines are the amplification curves for Ori (pink when the enzyme to degrade RNA:DNA hybrids (namely: DNase-XT) was used), and the blue lines are the amplification curves for Spike (red when the enzyme to degrade RNA:DNA hybrids was used).
N.B. Just to remind the reader, the farther along the x-axis the starting point of the line is, the less starting amount of DNA there is. The PCR cycle number usually ranges from 1 to 40–45 cycles where the x-axis shows the progression of amplification cycles and the y-axis shows the fluorescence signal intensity. So all in all, this represents the amount of amplified product detected in real time, increasing as amplification proceeds. The cycle-threshold (Ct) value is a single number derived from this x-y plot: it is the fractional cycle number (a point along the x-axis) where the amplification curve crosses a set fluorescence threshold line (a horizontal line on the y-axis). The bottom line is that lower values (earlier crossing) indicate higher starting amounts of target DNA (more template → signal rises faster).
We systematically added the DNase1 (first) and DNase-XT (second) enzymes to test whether or not there would be a change in Cts for Spike and Ori. When we added DNase1, there was a shift for Ori (non-hybrid-y region) to the right on x-axis meaning that it was digested by DNase1 and there was less starting DNA when it was applied. But take a look at the results of adding DNase1 to Spike. Not much of a shift at all, especially for the Moderna 025G23A, Pfizer GK0936 and Moderna AW4694B samples. Interesting. You can see this especially clearly in the Pfizer GK0936 sample.
But when we then added DNase-XT which degrades RNA:DNA hybrids, you guessed it! The Spike Ct shifted a lot which means that this DNA got degraded because the appropriate enzyme was used! Just look at that red line go! You’ll also notice that the Ori got degraded significantly by this enzyme and that is not an unexpected result as DNase-XT works well on dsDNA, ssDNA, RNA:DNA hybrids and chromatin.13 Makes you wonder why they didn’t just use it insteas od DNase1?
The main distinguishing feature of these results is that the DNase1 did not work on the Spike because it’s in a hybrid form. Those blue lines are effectively the same trajectory after treatment with DNase1; not so in the case of treatment with DNase-XT.
These are not small differences. They represent 100-1000X higher degradation of spike DNA.
Now, it bears repeating (I have said this multiple times in interviews) that Charles, Kevin and I did this work in 2 days (on Veteran’s Day no less - Charles is a Veteran) - as in, each of us did: 1. qPCR using 3 sets of primers (we also did SV40 in addition to Spike and Ori), 2. fluorometry (to quantify ALL of the DNA - not just the primer-focused DNA), and 3. Oxford Nanopore Sequencing to see the complete sequences popping out of these vials! Please read the paper to find out what we found there!
Hint:
Oxford Nanopore (ONT) sequencing further revealed numerous fragments exceeding 200 bp, including one read 5,284 bp in length that encompassed a large portion of the spike gene.
Do you believe that? Well done Charles. This is why it’s important to use techniques other than a KAN qPCR assay to look for DNA.
We did an incredible amount of work in 2 days and it was pretty much flawless because: 1. we had an excellent idea of the problem at hand before we even touched our pipettes, 2. we defined the problem precisely ahead of time in the form of 3 separate experiments to be done, 3. and we were led by Kevin who carefully took us through the protocols - he is very experienced and knows his own lab and pipettes. I want to give special thanks to Kevin for spending his off-time showing us these protocols and to Children’s Health Defense for partially helping with the cost of reagents. Also, thanks to Kevin for doing the dirty business of opening up the vacuum-sealed modRNA Pfizer/Moderna vials in the bio-hood: they are quite tricky to open since they have metal seals and one can get cut.
It’s still so wild to me that people got repeatedly injected with this stuff (and still are!) that should be treated and handled as biohazards. In the Moderna SOP for determination of residual DNA by qPCR in mRNA in the SAFETY section (7.0), it is advised that all people who handle these modRNA materials (for qPCR assay preparation) wear proper PPE.
But it’s ok to inject this stuff directly into you though. And babies.
All in all, I think the two main take home messages from our paper are:
That we proved (AGAIN) that there’s DNA in copious amounts (above the already too high EMA 10 ng/dose limit designed for naked DNA) in the Pfizer and Moderna COVID-19 “vaccine” vials, and
Why there’s DNA in copious amounts → easily anticipated RNA:DNA hybrids from improper choice of DNA enzyme for product cleaning during modRNA synthesis.
We also irrefutably argue why there was never going to be a way to actually show that there are boatloads of DNA in the vials: the manufacturers pre-designed the wrong assays for eventual use for detection.
That’s not ignorance - that’s malfeasance.
Please read the paper and share it with everyone. It’s not a long read and it’s written for understanding by the lay-person.
McKernan K, Rixey C, Rose J. RNA:DNA hybrids survive digestion in mRNA vaccine manufacturing. J Independent Med. 2026;2(1). doi:10.71189/JIM/2026/V02N01A04
Parr C. J. C., Wada S., Kotake K., Kameda S., Matsuura S., Sakashita S., et al. (2020). N 1-Methylpseudouridine Substitution Enhances the Performance of Synthetic mRNA Switches in Cells. Nucleic Acids Res. 48, e35. 10.1093/nar/gkaa070
Lenk R, Kleindienst W, Szabó GT, et al. Understanding the impact of in vitro transcription byproducts and contaminants. Front Mol Biosci. 2024;11:1426129. doi:10.3389/fmolb.2024.1426129
Sutton, D. H., Conn, G. L., Brown, T., and Lane, A. N. (1997). The dependence of DNase I activity on the conformation of oligodeoxynucleotides. Biochem. J. 321 (Pt 2), 481–486. doi:10.1042/bj3210481
Stephanie Seneff, Greg Nigh, Anthony M. Kyriakopoulos, Peter A. McCullough, Innate immune suppression by SARS-CoV-2 mRNA vaccinations: The role of G-quadruplexes, exosomes, and MicroRNAs, Food and Chemical Toxicology, Volume 164, 2022, 113008, ISSN 0278-6915, https://doi.org/10.1016/j.fct.2022.113008
S.A. Shabalina, N.A. Spiridonov, A. Kashina. Sounds of silence: synonymous nucleotides as a key to biological regulation and complexity. Nucleic Acids Res., 41 (4) (2013), pp. 2073-2094, 10.1093/nar/gks1205
McKernan K, Kyriakopoulos AM, McCullough PA (2021) Differences in Vaccine and SARS-CoV-2 Replication Derived mRNA: Implications for Cell Biology and Future Disease. doi: 10.31219/osf.io/bcsa6
For the record, they are not the excised introns themselves or their lariats.
Speicher, D. J., Rose, J., & McKernan, K. (2025). Quantification of residual plasmid DNA and SV40 promoter-enhancer sequences in Pfizer/BioNTech and Moderna modRNA COVID-19 vaccines from Ontario, Canada. Autoimmunity, 58(1). https://doi.org/10.1080/08916934.2025.2551517
Therapeutic Goods Administration. FOI 5286 - TGA KAN PCR documents. 2024. Page 5. Available from: https://www.tga.gov.au/sites/de- fault/files/2024-09/FOI%205286.PDF
There is a spike-specific qPCR assay developed or used by Pfizer/BioNTech (for BNT162b2/Comirnaty.
https://www.neb.com/en-us/products/m0570-dnase-i-xt
It is shocking and gives support to Mike Yeadon's arguments that 'mistakes were not made'
A crime , THE CRIME was committed. THEY knew EXACTLY what they did , no mistakes were made . Not one person has answered to this. 🤬Thank you Charles , Jessica and Kevin. 🩷🩷🩷