Applicability Target Sequences UniversalAll rAAV samples ITR2 primers: 5′-ggaacccctagtgatggagtt-3′ and5′-cggcctcagtgagcga-3′;probe: R-5′-cactccctctctgcgcgctcg-3′-Q. Semi-universalGroup of rAAV samples CMV promoter primers: 5′-ttcctacttggcagtacatctacg-3′ and5′-gtcaatggggtggagacttgg-3′;probe: R-5′-tgagtcaaaccgctatccacgccca-3′-Q;and other sequences. CAG promoter primers: 5′-ctgaccgcgttaatcccaca-3′ and5′-acaagccgtgattaaaccaaga-3′. CBA promoter […]
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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° […]
Why Are Transgenes Cloned into Lentiviral Vectors Without a Poly(A) Signal?
In lentiviral gene delivery systems, transgene expression cassettes are commonly cloned without a polyadenylation signal to prevent disruption of full-length lentiviral genomes on mRNA maturation in producer cells. Since lentivirus is […]
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 […]
Transfection Efficiency: What Makes Plasmid DNA Transfection Grade
Successful transient transfection relies on many factors, starting with a successful plasmid prep. To ensure high transfection efficiency, resulting DNA from your plasmid preps must be transfection grade. But what […]
Why Uncut Plasmid DNA on Agarose Gel Has 3 Bands
When uncut plasmid DNA is isolated and run on an agarose gel, you may observe two, three, or even four or more bands. Hopefully, the majority of your isolated DNA […]