- Low Endotoxin Level:
Animal-derived materials free, <0.005EU/μg, on request
- GMP-Like Manufacturing Pipeline:
Complete and detailed manufacturing documentation
- Strict Quality Control:
ISO9001/2015 quality management
- Fast procedure and on-time delivery
|Research Grade||Application Grade|
|Quantity||0.1 mg to gram level||1 mg to gram level|
|Turnaround time||Starting at 3 business days||Starting at 7 business days|
|Quality Control Methods||Basic quality control tests||Basic quality control tests plus Sterility assurance, Restriction enzyme analysis and Sequence verification|
|Standard Delivery Package Contains||Prepared plasmid DNA, Certificate of analysis (COA) and QC report|
Note：Synbio Technologies provides services upstream of plasmid preparation such as gene synthesis and subcloning into both commercial and custom vectors. This allows us to offer a unique approach to satisfy all our customer’s requests. Additionally, if you wish to provide the plasmid template, you will need to prepare over 100 ng DNA sample (diluted with ddH2O or TE buffer), colonies (fresh) or bacteria stored in glycerol.
1. The target plasmid DNA was transformed into bacteria cells and a monoclonal colony was selected to be cultured.
2. Restriction enzymes were used to verify the sequence of the mini-prepped plasmid.
3. Large volume bacteria culture or fermentation was performed to obtain a sufficient amount of cell paste.
4. The cell paste was lysed to release plasmid DNA. The crude lysis solution was then purified through several steps to dispose of impurities.
5. The bulk DNA solution was quality tested which included: restriction enzyme digestion, sterility tests and endotoxin level tests.
6. After passing all of the quality control tests, the bulk DNA was formulated, dispensed, labelled and finally delivered.
Quality Test Results:
1. Endotoxin Test: The negative control of endotoxin testing will not concrete, while the positive control will concrete. These samples were not concrete (Fig.1), proving that the endotoxin was successfully removed.
Fig. 1 Endotoxin testing, from left to right: negative control, positive control, sample, endotoxin standard 5UR/mg
2. Sterility Test No bacterial colony was detected after 48 h culturing in non-resistant medium (Fig.2).
Fig. 2 Bacterial testing result comparison
|Quality Control||Standard Range||Measurement Technique|
|Appearance||Clear, transparent||Visual inspection|
|A260/280 Analysis||1.80~2.00||UV spectrophotometer|
|Concentration||Up to 5mg/ml, default 1±0.05 mg/ml||UV spectrophotometer|
|Supercoil Percentage Analysis||>95%||Density scan on agarose gel|
|RNA Residue Analysis||Not detectable||Visual inspection on agarose gel|
|Genomic DNA Analysis||Not detectabl||Visual inspection on agarose gel|
|Sterility Assurance||No clone growth on LB plate > 48h||Incubate 0.1mg of product on LB plate for >48h|
|Restriction Enzyme Analysis (optional)||As expected, no other minor band||Incubate with enzyme and analyze on agarose gel|
|Sequence Verification (optional)||Correct||DNA Sequencing|
|Endotoxin Analysis (optional)||<0.005EU/μg||Endotoxin Analysis Assay|