Process and apparatus for forming a dry DNA transfer film, a transfer film product formed thereby and an analyzing process using the same

A process forming a plurality of spaced dry DNA dots on a glass substrate for testing purposes, said process comprising:

• placing liquid DNA solution within an array of spaced cup-shaped wells within a well plate;
• causing localized, momentary contact between an ink printing substrate and the liquid DNA within respective wells to create a corresponding series of spaced wet DNA spots on said ink printing substrate;
• drying said spots to form a dry DNA transfer media and placing said dry DNA transfer media in proximity to said glass substrate, with said dry DNA spots facing a surface of said glass substrate;
• and
• selectively, locally pressure impacting the opposite face of said dry DNA transfer media behind said dry DNA spots to cause mechanical transfer of dry DNA dots from said media spots to the facing surface of said glass plate.

A high throughput process for forming a dry DNA transfer film comprising:

• placing separated, replicated DNA genes solubilized in a solution within respective wells of a generally rigid well plate having within an upper surface thereof, a plurality of closely spaced wells;
• fixedly mounting a thin flexible resilient film against said upper surface and sealed to the upper surface of the well plate and effecting a rigid film plate assembly;
• inverting said rigid assembly to cause the DNA solution to physically, locally wet coat a surface of the film while preventing the DNA gene solution from running radially from one spot to another;
• reinverting said assembly to its initial position;
• removing the DNA gene solution spotted film slowly from the well plate;
• drying the DNA gene solution spot coating thereon and thereby forming a dry DNA transfer film capable of physical and chemical dry transfer of DNA to a test substrate.

A dry DNA transfer film formed by the process comprising:

• placing separated, replicated DNA genes solubilized in a solution within a plurality of spaced cup-shaped wells within a generally rigid well plate, with said cells opening to a top surface of said well plate;
• fixing a flexible resilient film having a roughened surface to said top surface of the well plate, with said roughened surface facing said well plate top surface;
• effecting a rigid film plate assembly;
• inverting said assembly to cause the DNA solution to physically, locally wet coat the roughened surface of the film, with the roughened surface causing the DNA gene solution to cling to the film as spaced dots corresponding to the well position while preventing the DNA gene solution from running radially from one spot to another;
• reinverting the assembly to its initial position;
• removing the DNA gene solution spotted film from the well plate;
• and
• drying the DNA gene solution spots to form a dry DNA transfer film capable of physical and chemical dry transfer of the DNA to a test substrate.

A dry form DNA analyzing process comprising the following steps:

• prefilling respective separated, replicated DNA genes solubilized in a solution within a plurality of upwardly open spaced wells within an upper surface of said rigid well plate;
• sealing a flexible resilient film at a predetermined position on the upper surface of said rigid well plate, with said film covering the prefilled wells;
• forming a fixed, sealed assembly between the well plate and the overlying thin flexible film;
• inverting the assembly to transfer DNA gene solution as spaced spots to the facing surface of the film over localized areas of said film defined by respective wells;
• reinverting the assembly;
• removing the film and drying the transferred DNA gene solution spots to thereby form a dry DNA gene transfer film;
• placing the dry DNA gene transfer film in a position facing a flat glass test substrate and momentary pressing the face of the dry DNA gene transfer film opposite that bearing said spots at spot locations, causing pieces of dry DNA to locally contact the test substrate such that portions of the dry DNA spots mechanically transfer from the transfer film to the test substrate by physical action and chemical attraction;
• sequentially repeating the last step at different localized dried DNA spot locations on the dry DNA gene transfer film to complete a dry DNA gene test dot array on the test substrate;
• and
• bathing the dots with two or more total genomic tag fluoresces to hybridize the tag to a particular gene of the array and optically scanning the array to identify dots containing fluorescently labeled DNA to determine the existence of mutants or lack of the same.

Apparatus for forming a dry DNA transfer media comprising:

• a table, said table including an elongated table base extending horizontally, a vertical riser extending upwardly from the base intermediate of ends of the base, a horizontal flat table top hinged at one end to said vertical riser, a fixed stop mounted upright to said base and underlying said table top remote from said first hinge, a cover overlying said table and hinged by a second hinge to said table top so as to extend parallel to said table top overlying the same in first position and being rotatable 1800 to an open position extending parallel to the table top and to the side of said center leg opposite that of said table top, a pivotable stop mounted to said base to the side of said center post opposite that of said fixed stop and underlying said cover when said cover is pivoted from said first position to said second position and being of a vertical height so as to maintain said cover horizontal when in said second position, the bottom surface of said cover being flat and including location pins for locating and fixing the position of said media on said bottom surface of said cover, means for positioning at least one well plate in fixed lateral and longitudinal positions on the upper surface of said table top at a position aligned with that of said transfer media, clamping means for fixing said cover in spaced overlying position on said table top, with said transfer media overlying said well plate, said well plate including a plurality of spaced upwardly open wells adapted to carry separated, replicated DNA genes solubilized in a solution within respective wells, said apparatus further comprising means for sealing the transfer media against the upper surface of said well plate and covering said upwardly open wells, whereby with said cover in said second position said transfer media may be fixed in a designated position on the bottom surface of said cover via location means, and said at least one well plate may be fixedly positioned at a predetermined position on the upper surface of said hinged table top, wherein by rotation of said cover 180° from said second position to said first position and by operation of said clamping means for fixing said cover in spaced overlying positions with respect to said table top, a fixed assembly may be achieved, whereupon subsequent rotation of said fixed assembly including said table top through a 180° from its position against the fixed stop in a direction towards said movable stop and by moving said movable stop to an inclined position on said table base, said assembly is inverted to cause the DNA solution to physically, locally wet coat the surface of said media creating spaced spots of DNA solution, whereupon the assembly may be pivoted about said first end through 180° to the extent of said table top contacting the fixed stop, whereupon the clamping means may be released permitting the cover to rotate from its first position to its second position to allow the media with the wet DNA coated spots to be removed from the table top and said spots to dry to thereby form said dry DNA transfer media.

Apparatus for forming a dry DNA transfer media capable of physical and chemical dry transfer of DNA to a test substrate, said apparatus comprising:

• a table, said table including a flat horizontal base, location means for locating a transfer media at a fixed position on an upper surface of said table top, riser means adjacent one end of said transfer media, a flat rectangular well plate holder pivoted at one end to an upper end of said riser means for pivoting of said well plate holder through an arc of at least 90° between a position overlying said transfer media and extending parallel thereto and a raised position, a locking pin and support means at an end of said well plate holder remote from said hinge and including a stop for supporting said well plate holder in a fixed position overlying said transfer media and parallel thereto, at least one elongated well plate hole within said well plate holder for receiving a well plate, said well plate including a plurality of spaced upwardly open cup-shaped wells for receiving separated, replicated DNA genes solubilized in a solution within respective wells, and means carried within said well for sealing off the upper open ends of said wells and for facilitating locally wet coating of the facing surface of said transfer media when said at least one well plate is inverted and inserted within a correspondingly sized opening within said well plate holder to the extent of said means closing off the open ends of said wells contacting the surface of said transfer media, whereupon release of said clamping means and rotating of said pivotable well plate holder to said raised position moves the well plates away from the wet DNA coated surface of said transfer media to permit removal of said transfer media from the upper surface of said table and to allow replacement thereof by a new uncoated transfer media and the process repeated.