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Description : EZgeneTM M13 isolation kit allows rapid and reliable isolation of high-quality M13 phage DNA from appropriate host strain. The system combines the reversible binding properties of our matrix with the speed and versatility of spin column technology to yield approximately 3 to 10 μg M13 phage DNA from 1-3 mL of phage supernatant. In this procedure, the infected bacterial culture is centrifuged and Buffer PC is used to precipitate the phage particle supernatant and then the sample is loaded to the column. The specially designed matrix will allow the phage particles to remain intact on the surface of the matrix. The phage particles are then lysed and bounded to the matrix with the addition of Buffer PB. The pure M13 DNA is eluted with Elution Buffer or ddH2O while unwanted proteins and other impurities are washed away by DNA Wash Buffer. All components are guaranteed for 12 months from the date of purchase when stored at 22-25°C. Buffer PB contains chaotropic salts, which may form reactive compounds when combines with bleach, Do not add bleach or acidic solutions directly to the preparation waste, ware gloves and protective eyewear when handling. Catalog# PD1316-00 PD1316-01 PD1316-02 Preps 4 50 250 DNA mini columns 4 50 250 2 mL collection tube 8 100 500 Buffer PB 1.5 mL 18 mL 80 mL Buffer PC 5 mL 80 mL 300 mL DNA Wash Buffer 2 mL 15 mL 3 x 24 mL Elution Buffer 1.5 mL 10 mL 25 mL User Manual 1 1 1 Exam this handbook and get familiar with each step. Prepare all components and have the necessary materials ready. Problems Possible reasons Suggestions Low DNA yield Incorrect host strain The host strain should carry the F'-episome that is essential for M13 infection. Low DNA yield Bacterial culture overgrown or not fresh. Do not grow culture for over 7 hours. Low DNA yield Low pH of elution buffer Make sure that the pH of elution buffer is in the range of 7.5-8.0. Low DNA yield Elution buffer added to the edge of membrane. Add the elution to the center of membrane and incubate for 10 minutes prior to elution. Low DNA yield Column clogged Use less than 3 mL of M13 phage supernatant per column. Avoid bacterial pellet during transfer. No DNA eluted Forgot to dilute the DNA was buffer with ethanol Add absolute ethanol as instructed. Genomic DNA contamination. Carry over of bacterial cells during transfer the supernatant to column Make sure not to carry over the pellet when transfer the supernatant to the column. Spin the samples twice. M13 DNA floats out of well while loading to agarose gel. Ethanol residues eluted with the M13 DNA Wash the column as instructed. Catalog # Product Name Preps Price $ PD1711-01 HP plasmid mini kit 50 60.00 PD1711-02 HP plasmid mini kit 250 260.00 PD1712-01 HP plasmid midi kit 10 69.00 PD1712-01 HP plasmid midi kit 25 160.00 PD1713-01 HP plasmid maxi kit 10 160.00 PD1713-01 HP plasmid maxi kit 25 380.00 PD1311-01 BAC/PAC isolation kit 50 90.00 PD1311-02 BAC/PAC isolation kit 250 400.00 PD1312-01 96 BAC/PAC isolation kit 4x96 450.00 PD1312-02 96 BAC/PAC isolation kit 20x96 2100.00 PD1313-01 ezFilter 96 BAC/PAC isolation kit 4x96 500.00 PD1313-02 ezFilter 96 BAC/PAC isolation kit 20x96 2300.00 PD1316-01 M13 isolation kit 50 70.00 PD1316-01 M13 isolation kit 250 280.00 PD1318-01 96 ezFilter M13 isolation kit 4x96 120.00 PD1318-02 96 ezFilter M13 isolation kit 20x96 380.00Introduction
Storage and Stability
Safety Information
Kit content
Before Starting
Trouble shooting Guide
Related EZgeneTM Products
- Model: PD1316-02
- Manufactured by: Biomiga
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