METHODS AND DEVICES FOR NUCLEIC ACID PURIFICATION

摘要

The invention provides pipette tip columns and automated methods for the purification of nucleic acids such as plasmids from unclarified cell lysates containing cell debris as well as methods for making and using such columns. The columns typically include a bed of medium positioned in the pipette tip column, above a bottom frit and with an optional top frit.

主权项

1. An automated method for purifying genomic DNA from an unclarified cell lysate with a pipette tip column, the method comprising:
a) providing a multi-well plate, wherein a plurality of wells in the multi-well plate contain unclarified cell lysates, wherein each unclarified lysate is comprised of genomic DNA, liquid and particulates; b) providing a plurality of pipette tip columns arranged in a 9 mm center-to-center format, wherein each pipette tip column is comprised of
i) a column body having an open upper end,ii) an open lower end,iii) an open channel between the open upper end and the open lower end,iv) a bottom frit extending across the open lower end, andv) a solid phase positioned inside the open channel and above the bottom frit; c) passing each unclarified lysates through a pipette tip column; d) passing a wash solution through each pipette tip column; e) transferring the pipette tip columns into a custom 9 mm center-to-center block, wherein the block is comprised of channels, wherein the channels completely surround each individual pipette tip column, wherein the block is designed to apply vacuum evenly through all the pipette tip columns; and f) applying software-controlled vacuum through the block, wherein the vacuum is applied evenly through each pipette tip column.