Pour la purification rapide de l'ADN total des bactériophages.
Détails
Supporting Data
Figure 1. Effective Host Genomic DNA Removal without Reducing Phage DNA Yield.
Total DNA was isolated from four enriched phage cultures using Norgen's Phage DNA Isolation Kit. A DNase I pre-treatment was performed prior to adding the provided Lysis Buffer. Briefly, 20 units of DNase I was added to 1 mL of enriched phage culture and the mixture was incubated at room temperature for 20 minutes. After the DNAase I treatment the procedure was followed. As a control, DNA was isolated from aliquots of the same 4 cultures using Norgen’s Phage DNA Isolation Kit without performing the DNase I treatment. For DNA analysis 10 µL of each 50 µL elution was loaded onto a 1X TAE agarose gel. As it can be seen, the phage DNA was safely protected from the DNase I treatment by its coat protein, while the host genomic DNA was efficiently degraded by the DNase I. Thus the DNase I pre-treatment resulted in less host gDNA contamination in the final phage elution without influencing the total phage DNA yield. Lane M is Norgen's Highranger 1 kb DNA Ladder (Cat. 11900)
Figure 2. Optional Proteinase K Treatment Improves DNA Yield for Certain Phage Strains.
Total DNA was isolated with and without the optional Proteinase K treatment using Norgen's Phage DNA Isolation Kit. Briefly, 4 µL of Proteinase K (20 mg/mL) was added to 1 mL of enriched phage culture and incubated at 55°C for 15 minutes with the phage Lysis Buffer. After the Proteinase K treatment the procedure was followed. As a control, DNA was isolated from aliquots of the same 8 cultures using Norgen's Phage DNA Isolation Kit without performing the Proteinase K treatment. For DNA analysis 10 µL of each 50 µL elution was loaded onto a 1X TAE agarose gel and the yield of DNA was compared from the eight different phage types (lane 1 to 8). As it can be seen, the optional treatment of Proteinase K improved the phage DNA yield in Lanes 2, 5 and 6 dramatically. Lane M is Norgen's Highranger 1 kb DNA Ladder (Cat. 11900)
Données justificatives
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Spécifications du kit
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Capacité de liaison de la colonne
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50 µg
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Volume maximal de chargement de la colonne
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650 µL
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Taille de l'ADN purifié
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Toutes les tailles
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| Quantité maximale de matériel de départ |
1 x 1010 pfu/mL de phages enrichis
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| Rendement moyen* |
3 à 15 µg d'ADN à partir de 106 à 1010 pfu/mL
de phages enrichis |
| Temps nécessaire pour effectuer 10 purifications |
45 minutes
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* Les rendements moyens varient en fonction des conditions utilisées et du stade de développement.
Conditions de stockage et stabilité du produit
Toutes les solutions doivent être maintenues hermétiquement fermées et conservées à température ambiante. Ce kit est stable pendant un an à compter de la date d'expédition.