Description : Introduction Key to the kit is our proprietary DNA binding systems that allow the high efficient binding of DNA to our ezBindTM matrix while proteins and other contaminates are removed under certain optimal conditions. Nucleic acids are easily eluted with sterile water or Elution Buffer. The purified DNA is ready for downstream applications such as cloning/subcloning, RFLP, sequencing, and transfection of HEK293 cells. This kit is especially designed for purifying plasmid DNA from endA+ strains such as HB101, JM101, TG1 or their derived strains. Important Notes The yield of plasmid DNA depends on the origin of the replication and the size of the plasmid. The protocols are optimized for high copy number plasmid purification. For low copy number plasmids, both the culture volume and the buffer volume need to be scaled up 3 to 5 times. Please contact our customer service for further information and reference the table below for the commonly used plasmids, Plasmid Origin High copy Low copy pACYC P15A 10-12 pSC101 pSC101 5 pSuperCos pMB1 10-20 pBR322 pMB1 15-20 pUC Muted pMB1 500-700 pGEMR Muted pMB1 300-400 pBluescriptR ColE1 300-500 Storage and Stability Buffer A1 should be stored at 4 ℃ once RNase A is added. All other materials can be stored at room temperature. The Guaranteed shelf life is 18 months from the date of purchase. Kit Contents Catalog Number PD1711-00 PD1711-01 PD1711-02 Preps 4 50 250 ezBindTM Columns 4 50 250 Buffer A1 1.2 mL 15 mL 70 mL Buffer B1 1.2 mL 15 mL 70 mL Buffer N1* 1.5 mL 20 mL 90 mL DNA Wash Buffer** 2 mL 15 mL 3 x 24 mL Buffer KB 2.4 mL 30 mL 135 mL EndoClean Buffer (Optional) 100 uL 1 mL (PD1214) 5 mL (PD1214) RNase A 1.2 mg 1.5 mg 7.0 mg *Buffer N1 contains chaotropic salts, wear gloves and protective eyewear when handling. **Add 8mL(PD1711-00)60 mL (PD1711-01) or 96 mL (PD1711-02) 96-100% ethanol to DNA Wash Buffer before use. The Final ethanol is 80% (v/v) Before Starting Prepare all components and get all necessary materials ready by examining this instruction booklet and become familiar with each steps. Important Materials supplied by users Trouble Shooting Guide Low Yield Poor Cell lysis. Low Yield Bacterial culture overgrown or not fresh. Grow bacterial 12-16 hours. Spin down cultures and store the pellet at -20 ℃ if the culture is not purified the same day. Do not store culture at 4 ℃ over night. Low Yield Low copy-number plasmid. Increase culture volume (up to 10mL for Miniprep). Scale up the volume of buffers accordingly. No DNA Plasmid lost in Host E.coli Prepare fresh culture. Genomic DNA contamination Over-time incubation after adding Buffer B1. Do not vortex or mix aggressively after adding buffer B1. Do not incubate more than 5 minutes after adding Buffer B1. RNA contamination RNase A not added to Buffer A1. Add RNase A to Buffer A1. Plasmid DNA floats out of wells while running in agarose gel, DNA doesn't freeze or smell of ethanol Ethanol traces not completely removed from column. Make sure that no ethanol residual remaining in the silicon membrane before elute the plasmid DNA. Re-centrifuge or vacuum again if necessary.
- Model: PD1711-02
- Manufactured by: Biomiga