Escherichia coli (E.coli), commonly known as Escherichia coli, is a Gram-negative short bacillus with a size of 0.5×1.3 micrometers. It is a type of Gram-negative short bacillus with flagella all over its body, rounded ends, motility, and no spores.
The insulin approved by regulatory authorities for the treatment of diabetes in 1982 made Escherichia coli the first expression host for the production of biological drugs. The Escherichia coli expression system is highly favored for its advantages such as low cost, high yield and easy operation, and is thus often used as the preferred expression system for recombinant proteins (such as recombinant cytokines, recombinant hormones, recombinant enzymes, recombinant vaccines, etc.).
Zhuhai Baorui Biotechnology Co., Ltd. has developed the E.coli Total RNA Residue Detection Kit (PCR-fluorescence probe method), which can be used in conjunction with Baorui Biotechnology's DNA Removal Kit and Nucleic Acid Extraction or Purification Kit II (magnetic bead method), providing a complete solution from extraction to detection.
Both the "Technical Guidelines for Pharmaceutical Research and Evaluation of In Vivo Gene Therapy Products (Trial)" and the "Technical Guidelines for Pharmaceutical Research and Evaluation of In Vitro Gene Modification Systems (Trial)" propose that process-related impurities are mainly introduced by the production process, such as host cell proteins, host cell DNA, host cell RNA, packaging plasmids, and endotoxins from viruses and bacteria used for packaging. It is necessary to establish reasonable quality standards and conduct release inspections.
The E.coli Total RNA Residue Detection Kit from Baorui Biotechnology employs the PCR-fluorescence probe method. It is designed with primer probes specifically for the 23S conserved sequence. The project uses ready-to-use calibrators to detect the residual amount of total RNA of the target gene copy number in the sample.
The reagent has excellent performance /
1. Quantification limit: 0.2 fg/μL.
2. Repeatability (intra-analytical precision), intermediate precision (inter-instrument, inter-day, inter-personnel, inter-reagent batch) and reproducibility (total imprecision), with a quantification limit CV≤20%, and a CV≤15% for medium and high concentrations.Seven genomes, including Vero, Mycobacterium, HEK293, CHO, MDCK, BHK21 and SP2/0, do not interfere with this kit.
/ Convenient and fast operation /1. Extraction is only requiredThree steps; The amplification time is only 90 minutes.
2. Calibrators do not need to be diluted, and the finished products can be assembled conveniently and quickly.
3. The fastest time is 2 hoursThe entire inspection can then be completed.
/ Anti-pollution design /
The UDG enzyme anti-contamination system is adopted to effectively avoid the risk of contamination of amplification products.
Freeze-thaw stability: The performance of the kit remains unaffected after repeated freeze-thaw cycles of 3, 5, 7, and 10 times.
2. Instrument applicability: The performance is not affected on different models of equipment such as ABI 7500, ABl QuantStudio™ 5, Bio-Rad CFX96, SLAN-96P, SLAN-96S, and Roche Light cycler 480 Ⅱ.
References
"Technical Guidelines for Pharmaceutical Research and Evaluation of Gene Therapy Products in Vivo (Trial)"
2. Technical Guidelines for Pharmaceutical Research and Evaluation of In Vitro Gene Modification Systems (Trial)
3.《2016 Pharmacopeial PF 42(5) Forum Proposed USP General Chapter <509> Residual DNA Testing》
