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Quantification for residual host cell DNA contamination in the rAAV prep via qPCR or ddPCR analysis targeting highly repetitive genome sequences

DNA Impurity Target Sequences Plasmid KanR primers: 5′-gggcgcccggttctttttgtc-3′ and5′-gccagtcccttcccgcttcagtg-3′. AmpR primers: 5′-cgcgccacatagcagaactt-3′ and5′-cgccccgaagaacgttt-3′;probe: R-5′-aaaagtgctcatcattg-3′-Q;and other sequences. Adenovirus E2A primers: 5′-ttgctgaaacccaccatttg-3′ and5′-tcgtggacagcgaggaaga-3′;probe: R-5′-cgccacatcttctct-3′-Q. Adenovirus E4 primers: 5′-tcggcgcactccgtaca-3′ and5′-cgcgggtctctgtctcaaaa-3′;probe: R-5′-tagggatcgcctacctc-3′-Q. HSV UL23 […]

How to protect RNA from degradation?

To protect RNA from degradation, you can:  RNA is prone to enzymatic degradation and hydrolysis. Even at low temperatures, RNA can still be reactive, and some ribozymes can be enhanced by […]

What is the main reason for RNA being unstable?

Unlike DNA, RNA is usually single-stranded. Additionally, RNA contains ribose sugars rather than deoxyribose sugars, which makes RNA more unstable and more prone to degradation. RNA is generally stable at –80° […]

Potential Reasons for Lack of Supercoiled DNA

Conclusion In conclusion, the absence of apparent supercoiling in plasmid DNA preparations can result from a variety of factors related to DNA extraction, purification, handling, and analysis. By systematically addressing […]