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USER INSTRUCTIONS Standard delivery* One vial containing approximate 4 μg of lyophilized plasmid DNA with your insert. Applicable to all gene synthesis, mutagenesis, and vector construction services Storage Conditions Plasmids are delivered in vials with red or blue caps; Store the vials at 4°C for short-term and frequent use, or at -20°C for long-term storage. Material Instructions Keep the vial sealed until ready to use. Centrifuge at 6,000 x g for 1 minute at 4°C. Open the vial and add 20 μl of sterilized water to dissolve the DNA. Close the lid and vortex the vial for 1 minute. If necessary, heat the solution at 50°C for 15 minutes to dissolve the DNA. Transformation of the plasmid DNA can be performed directly after completion of the steps above. Verify the plasmid sequence after each subcloning and transformation step. PCR products & Scale-up plasmid delivery Storage Conditions PCR products & Scale-up plasmids are delivered in vials with red or blue caps; Store the vials at 4°C for short-term, frequent use, or at -20°C for long-term storage. Material Instructions If the plasmid DNA is lyophilized: Keep the vial sealed until ready to use. Centrifuge at 6,000 x g for 1 minute at 4°C. Open the vial and add 20 μl of sterilized water to dissolve the DNA. Close the lid and vortex the vial for 1 minute. If necessary, heat the solution at 50°C for 15 minutes to dissolve the DNA. If the plasmid DNA is dissolved in solution: Centrifuge the tube briefly before opening. The concentration of the plasmid DNA is indicated in the COA file. If the concentration is > 3 mg/ml, the solution may be thick and sticky. Vortex the solution briefly and heat at 50°C for 15 minutes if necessary to dissolve the DNA. Make dilutions when appropriate before use. Transformation of the plasmid DNA can be performed directly after completion of the steps above. Verify the plasmid sequence after each subcloning and transformation step. * a. For regular gene synthesis orders, synthetic gene is delivered in GenScript’s standard vector pUC57; please find the sequence, vector map, and restriction site information from the following link: http://www.genscript.com/vector/SD1176-pUC57_plasmid_DNA.html b. For gene synthesis orders combined with subcloning services, mutagenesis, or custom cloning and siRNA/miRNA orders, custom genes will be cloned into the customer's specified plasmid. The plasmid name is indicated in COA and on the vial label Glycerol stock Storage Conditions Glycerol stocks are delivered in vials with green caps; Store the vials at 4°C for short-term and frequent use, or at -80°C for long-term storage. Material Instructions The glycerol stock contains 15% glycerol, the stock host is E. coli (TOP10 strain), or as labeled on the vial. It can be used directly according to your requirements, pay attention to avoid contamination. Avoid repeated thawing and re-freezing of the glycerol stocks since it may reduce the viability of the bacteria. Transformation of the plasmid DNA can be performed directly after completion of the steps above. Please verify the plasmid sequence after each subcloning and transformation step. Bac-stab Storage Conditions Bac-stabs are delivered in vials with green caps. Store the vials at 4°C; do not store at -20°C. Not suitable for long-term storage. Material Instructions The gene product is transformed into E. coli TOP10 cells or as labeled on the vial. Cells can be cultured directly from the bac-stab. Add 500 μl of LB to the bac-stab vial in a clean, sterile hood, and incubate it at 37°C for 30 minutes. Streak the bacterium onto an LB medium plate. Incubate the plate at 37°C overnight. Pick up a single clone to culture and to perform a plasmid prep on. Transformation of the plasmid DNA can be performed directly after completion of the steps above. Verify the plasmid sequence after each subcloning and transformation step. Files instructions Please view contig printout to verify if the sequence is 100% accurate. If any mismatches are found, contact our customer service immediately. Note, green account users will not receive printout of contigs; you may login to your online account to download them. To view sequence chromatograms, login to your account via https://www.genscript.com/ssl-bin/myaccount and click “My Orders” on the left navigation, then click Order ID & scroll down to view the chromatogram data and construct map. If you forgot your password, click “Forget your password?” on your login page and follow the instructions on the screen. Sequencing chromatogram (trace) data can be viewed by using Chromas (PC) or Editview (Mac). Sequence alignment files can be opened by DNASTAR® SeqMan. Construct map (for gene synthesis, mutagenesis, and cloning service) can be opened by Adobe PDF Reader. www.genscript.com 860 Centennial Ave., Piscataway, NJ 08854, USA | Toll-Free: 1-877-436-7274 | Tel: 1-732-885-9188 | Fax: 1-732-210-0262 | Email: [email protected] All Rights Reserved. All content described by GenScript is copyright of GenScript Corporation unless specifically identified otherwise. This includes all imagery, text and programmatic computer code.