The pLV3-CMV-H4C1(human)-3×HA-CopGFP-Puro plasmid is a plasmid suitable for mammalian cells and can also be used for lentiviral packaging.
Instructions
Ⅰ. Plasmid dry powder (stored at minus 20 degrees, shelf life 90 days, transportation at room temperature)
1. After receiving the dry powder plasmid, please centrifuge at 5000rpm for 1min, and then add 20μl of sterile water to dissolve the plasmid;
2. Thaw the corresponding competent on an ice box, take 2 μl of plasmid and add it to 100 μl of competent for 30min;
3. 42 degrees heat shock for 60s, ice bath for 2min, add 900μl of antibiotic-free LB medium, and culture at 180rpm shaking for 45min;
4. Centrifuge at 6000rpm for 3min, leave 100μl of supernatant to resuspend, and then coat it on the corresponding resistant LB plate;
5.After inverting overnight, if there are too many colonies, the plasmid will be diluted for transformation, and if there are no colonies, 10ul plasmid will be taken for transformation;
6. Pick single colonies into the corresponding resistant LB liquid medium overnight shaking culture, and extract the plasmid as needed.
Ⅱ. Liquid plasmid (minus 20 degrees storage, shelf life 90 days, ice pack transportation).
1. After receiving the liquid plasmid, please shake the preservation tube so that the liquid plasmid does not adsorb on the tube wall;
2.Take an appropriate amount of plasmid according to the experimental needs, and store the rest in a minus 20 degree freezer.
Ⅲ. Glycerol strains (stored at minus 80 degrees, shelf life of 90 days, ice pack transportation).
1. After receiving the glycerol strain, please shake the preservation tube so that the bacterial solution does not adsorb on the tube wall;
2. Use the inoculation loop to mark the line in four areas on the solid plate, and add the corresponding resistance if there is resistance;
3. Inverted culture in constant temperature incubator for 12~16h;
4. Be sure to pick a single colony and inoculate it in the liquid medium, and add the corresponding resistance if there is resistance;
5. Shake the shaker and culture overnight, and culture the strains according to the needs of the experiment.
产品声明
For Research Use Only All products available through the MiaoLing Plasmid Platform are intended for research use only. They are not intended for clinical trials or for use in humans, including oral administration, injection, or therapeutic use in vivo or ex vivo.
Plasmid Sequence and Experimental Performance Plasmid products are provided and sold based on the corresponding reference/template plasmid. MiaoLing guarantees the accuracy of the specified gene sequence based on our sequencing results, but does not guarantee downstream experimental performance, including gene expression, transfection efficiency, protein expression, or other experimental outcomes.
Sequence Verification of Certain Vector Products For certain vector products, even the original manufacturer may not have fully verified the complete plasmid sequence. If the sequence of the key functional regions has been confirmed to be correct, the product will be considered a correct and usable vector for research purposes.
After-Sales Period for Small-Pack Plasmid Products The after-sales service period for small-pack plasmid products is three months from the date of receipt. Customers are advised to transform and process the plasmid promptly after receipt. After the three-month period, a new purchase may be required for further support or replacement.
Research-Related Risks and Limitation of Liability Scientific research is inherently exploratory and involves a significant degree of uncertainty. To the maximum extent permitted by applicable law, MiaoLing shall not be liable for any additional economic losses, indirect losses, or other liabilities arising from the use of our products or from research outcomes.
Citation in Chinese Publications For publications in Chinese, please cite the product as follows: 产品名称（产品货号）由淼灵质粒平台提供。
Citation in English Publications For publications in English, please cite the product as follows: “Product Name (Product Item Number) was obtained from MiaoLing Plasmid, China.”
