Lid for pcr vessel comprising probes permitting pcr amplification and detection of the pcr product by hybridisation without opening the pcr vessel

Abstract

The present invention proposes a simple and effective lid being part of a device having multiwells for performing simultaneous amplifications by PCR and detections of multiple target molecules on unlabeled capture molecules immobilized on the lid.

1 . A sealable lid for a PCR multiwell plate provided with at least one defined area, said defined area being configured to be mated to at least one well of the multiwell plate, at least one of said defined area being pretreated to be provided with unlabeled capture molecule. 
     
     
         2 . The lid of  claim 1 , wherein at least one defined area bears an immobilized unlabeled capture molecule. 
     
     
         3 . The lid of  claim 2 , wherein the capture molecule is a polynucleotide. 
     
     
         4 . The lid of  claim 2 , wherein different defined areas of the same lid have fixed the same capture molecules. 
     
     
         5  The lid of  claim 1 , wherein the lid contains at least 2, preferably at least 6, more preferably at least 24, still more preferably at least 96 defined areas. 
     
     
         6 . The lid of  claim 1 , wherein the multiwell plate has at least 2, preferably at least 8, more preferably at least 24, still more preferably at least 96, even more preferably at least 384 wells. 
     
     
         7 . The lid of  claim 1 , wherein the wells of the multiwell plate which are separated from each other by a pitch of 4.5 mm or a multiple thereof. 
     
     
         8 . The lid of  claim 1 , wherein the measurement of the amplified target nucleotide molecules is performed on the lid of the multiwell plate after hybridization on capture molecules. 
     
     
         9 . The lid of  claim 1 , wherein the lid is used for real-time PCR measurement. 
     
     
         10 . The lid of  claim 1 , wherein the lid is used for end point PCR measurement. 
     
     
         11 . The lid of  claim 2 , wherein the defined area has fixed upon its surface at least 4 different capture molecules being physically separated in compartments. 
     
     
         12 . The lid of  claim 2 , wherein the defined area has fixed upon its surface a micro-array comprising at least 5 different capture molecules being immobilized in specifically localized areas of said defined area. 
     
     
         13 . The lid of  claim 12 , wherein the micro-array comprises more than 10 different capture molecules, preferably more than 20 capture molecules, more preferably more than 50 capture molecules. 
     
     
         14 . The lid of  claim 1 , wherein the defined area has aldehyde groups fixed upon their surface. 
     
     
         15 . The lid of  claim 14 , wherein the defined area is divided in at least 4 compartments. 
     
     
         16 . The lid of  claim 15 , wherein the defined area is used to fix capture molecules terminated by a primary amino group. 
     
     
         17 . The lid of  claim 1 , wherein the lid is maintained flat at temperature higher than 85° C. 
     
     
         18 . The lid of  claim 1 , wherein the defined area has a flatness tolerance of less than 100 microns and better of less than 25 microns. 
     
     
         19 . The lid of  claim 1 , wherein the light transmittance of the lid at the wavelength used for the detection is higher than 80%, preferably higher than 90%. 
     
     
         20 . The lid of  claim 1 , wherein the lid comprises or is made of a material selected from the group consisting of glass, metal and polymer. 
     
     
         21 . The lid of  claim 20 , wherein the polymer is selected from the group consisting of: polycarbonate (PC), polyethylene (PE), Cycloolefin copolymer (COC), cyclic olefin polymer (COP) or a mixture thereof. 
     
     
         22 . The lid of  claim 1 , wherein the defined area is configured to be mated to at least one well of the multiwell plate by means of a sealable material. 
     
     
         23 . The lid of  claim 1 , wherein the defined area is configured to be mated to at least one well of the multiwell plate by means of an adaptor comprising a sealable material. 
     
     
         24 . The lid of  claim 22 , wherein the sealable material is made of a soft polymer or a glue. 
     
     
         25 . A sealable lid ( 1 ) for a PCR multiwell plate provided with at least one defined area, said defined area being configured to be mated to at least four wells of the multiwell plate, at least one of said defined area being pretreated to be provided with unlabeled capture molecule. 
     
     
         26 . The lid of  claim 25 , wherein the lid contains at least 2, preferably at least 6, more preferably at least 24, still more preferably at least 96 defined areas. 
     
     
         27 . The lid of  claim 25 , wherein the multiwell plate has at least 2, preferably at least 8, more preferably at least 24, still more preferably at least 96, even more preferably at least 384 wells. 
     
     
         28 . The lid of  claim 25 , wherein the wells of the multiwell plate which are separated from each other by a pitch of 4.5 or 9 mm. 
     
     
         29 . The lid of  claim 25 , wherein the lid is used for end point PCR measurement. 
     
     
         30 . The lid of  claim 25 , wherein the defined area has aldehyde groups fixed upon their surface. 
     
     
         31 . The lid of  claim 25 , wherein the defined area is divided in at least 4 compartments. 
     
     
         32 . The lid of  claim 25 , wherein the defined area is used to fix capture molecules terminated by a primary amino group. 
     
     
         33 . The lid of  claim 25 , wherein the lid is maintained flat at temperature higher than 85° C. 
     
     
         34 . The lid of  claim 25 , wherein the defined area has a flatness tolerance less than 100 microns and better less than 25 microns. 
     
     
         35 . The lid of  claim 25 , wherein the light transmittance of the lid at the wavelength used for the detection is higher than 80% preferably higher than 90%. 
     
     
         36 . The lid of  claim 25 , wherein the lid comprises or is made of a material selected from the group consisting of glass, metal, polymer. 
     
     
         37 . The lid of  claim 36 , wherein the polymer is selected from the group consisting of: polycarbonate (PC), polyethylene (PE), Cycloolefin copolymer (COC), cyclic olefin polymer (COP) or a mixture thereof. 
     
     
         38 . The lid of  claim 25 , wherein the defined area is configured to be mated to at least one well of the multiwell plate by means of a sealable material. 
     
     
         39 . The lid of  claim 25 , wherein the defined area is configured to be mated to at least one well of the multiwell plate by means of an adaptor comprising a sealable material. 
     
     
         40 . The lid of  claim 38 , wherein the sealable material is made of a soft polymer or a glue.