Description :

Introduction

The 96-well BAC or PAC DNA Isolation Kit is designed for rapid high-throughput purification of BACs, PACs, and P1s from small volume of E. Coli.bacterial cultures grown and processed in a 96-well plate format. Process may be performed by using either a centrifugation or full vacuum protocol. One 96-well plate can be processed manually within 60 minutes and two plate in less than  80 minutes.  The procedure has been developed and tested using a variety of low copy cosmids, BACs, PACs, P1s , and E. coli strains. This kit can also be used for high copy plasmid isolation.

The 96-well  Fastfilter BAC/PAC DNA Isolation procedure is based on modified alkaline lysis procedure in which the bacterial cells are lysed in the presence of RNase A, After the naturalization state, the cell lysates are cleared by filtration using 96-well Lysate Clearance Plate. The liberated BAC, PAC, P1, or plasmid is trapped on to the membrane in the  96-well  DNA Plate with a propriety technology. After two quick wash step, the purified BAC, PAC or plasmid DNA can be eluted with low Elution Buffer or water. The eluted BAC, PAC or plasmid can be directly used in downstream applications.

Materials supplied by user

•  Centrifuge with swing-bucket rotor (4,000 x g).
•  Please disinfect 96-well 2.2 mL plates before use.
•  Vacuum pump capable of achieving 300-400 mbar.
•  Standard vacuum manifold.
•  Oven or incubator preset to 70 ^oC.

Storage and Stability

All components are guaranteed for 24 months from the date of purchase. The Buffer A1/RNase A should be stored at 4^oC.

Kit Contents

Before Starting

Exam this handbook and get familiar with each step. Prepare all components and have the necessary materials ready.

•  Briefly spin down the RNase A vial and add the RNase A to Buffer X1.
•  Dilute DNA Wash Buffer as follows:

PD1312-00: Add 200 mL 96-100% ethanol to each bottle before use.

PD1312-01: Add 400 mL 96-100% ethanol to each bottle before use.

PD1312-02: Add 500 mL 96-100% ethanol to each bottle before use.

•  Dilute Binding Buffer as follows:

PD1312-00: Add 32 mL 96-100% isopropanol to each bottle before use.

PD1312-01: Add 128 mL 96-100% isopropanol to each bottle before use.

PD1312-02: Add 320 mL 96-100% isopropanol to each bottle before use.