The OmpA signal peptide is a 21-amino acid sequence that targets fused proteins to the Sec secretion pathway in E. coli. It can be used to increase protein yield and simplify purification […]
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Isopycnic Centrifugation
In isopycnic separation, also called buoyant or equilibrium separation, particles are separated solely on the basis of their density. Particle size only affects the rate at which particles move until […]
High-cell-density cultivation (HCDC)
High-cell-density cultivation (HCDC) is a process that improves the formation of microbial biomass and products. It can be used for microorganisms such as bacteria, archaea, and yeasts. HCDC can make the fermentation process […]
Is Kozak sequence essential for transgene expression in lentivirus?
Yes, a Kozak sequence is essential for efficient translation and transgene expression in lentiviruses: In addition to the Kozak sequence, a woodchuck post-transcriptional regulatory element (WPRE) can also be added […]
Primers and probes used for rAAV quantitative analysis
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 […]
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 […]