Ready-to-use electroporation cuvette including frozen electrocompetent cells

Abstract

A ready-to-use electroporation cuvette is provided that includes a cuvette, first and second electrodes positioned within the cuvette and electroporation competent cells frozen in a suspension solution within the cuvette, wherein the electroporation cuvette is configured to permit electroporation of the cells when the cells are thawed. The electroporation cuvette may be sealed with a cap that may be color coded to aid the user.

1 - 43 . (canceled) 
     
     
         44 . An electroporation cuvette with an internal structure comprising:
 first and second electrodes positioned at a first fixed distance apart within the electroporation cuvette,
 wherein each of first and second electrodes is in the form of an I-beam, each I-beam comprising (i) inner, and (ii) outer plates that are connected by at least one perpendicular interconnecting plate, the length of which determines, between the inner and outer plates, a second fixed distance; 
   a first space formed between the first and second electrodes;
 wherein the length of at least one of the interconnecting plates of the first and second electrodes may be varied to enable a change in the first fixed distance between the first and second electrodes. 
   
     
     
         45 . The electroporation cuvette of  claim 44  wherein the first and second electrodes positioned within the electroporation cuvette are held in position by ledges or walls within the electroporation cuvette. 
     
     
         46 . The electroporation cuvette of  claim 44  wherein at least one of the first and second electrodes is placed against and fixed to one of the inside walls of the electroporation cuvette. 
     
     
         47 . The electroporation cuvette of  claim 44  wherein the first and second electrodes are positioned parallel to each other. 
     
     
         48 . The electroporation cuvette of  claim 45  wherein the first and second electrodes are positioned parallel to each other. 
     
     
         49 . The electroporation cuvette of  claim 46  wherein the first and second electrodes are positioned parallel to each other. 
     
     
         50 . The electroporation cuvette of  claim 44  further comprising a second space formed in the upper portion of the electroporation cuvette above the first space, wherein the structure defining the boundaries of the second space comprises a wedge-shaped blend between the upper walls of the electroporation cuvette and the first and second electrodes, wherein the wedge-shaped blend provides a transition from the second space into the first space to facilitate the flow of liquids into the first space. 
     
     
         51 . The electroporation cuvette of  claim 50  wherein the structure defining the boundaries of the second space comprises one or more ledges to hold first and second electrodes in a fixed position during electroporation. 
     
     
         52 . The electroporation cuvette of  claim 44  wherein at least one of the first and second electrodes is placed against a one of the lower inside-walls of the electroporation cuvette. 
     
     
         53 . The electroporation cuvette of  claim 52 , wherein the first and second electrodes are positioned parallel to each other. 
     
     
         54 . The electroporation cuvette of  claim 44  wherein the inner and outer plates of the I-beam electrodes have sufficient structural rigidity to retain their shape, meaning that the fixed distance between the electrodes is maintained during freezing and thawing processes. 
     
     
         55 . The electroporation cuvette of  claim 44  wherein at least one of the first and second electrodes comprises a metal. 
     
     
         56 . The electroporation cuvette of  claim 44  wherein at least one of the first and second electrodes comprises aluminum. 
     
     
         57 . The electroporation cuvette of  claim 44  wherein the first electrode is fixed to the inside wall of the cuvette and the second electrode is placed against the inside wall opposite the first electrode of the cuvette, wherein the electrodes are positioned at a fixed distance apart. 
     
     
         58 . The electroporation cuvette any of  claim 50  wherein the structure defining the boundaries of the second space comprises one or more ledges to hold the second electrode in a position during electroporation. 
     
     
         59 . An electroporation cuvette with a structure comprising;
 first and second electrodes (i) positioned at a fixed distance apart, and (ii) fused to two of the directly opposing, lower walls of the electroporation cuvette;   wherein each of first and second electrodes is in the form of an I-beam, comprising:   parallel (i) inner, and (ii) outer plates that are connected by at least one perpendicular and variable-length interconnecting plate, the length of which determines a second fixed distance between the inner and outer plates of each of first and second electrodes;   wherein the directly opposing, lower walls of the electroporation cuvette pass through a second fixed distance between the inner and outer plates of each of the first and second electrodes;   wherein the outer plate of each of the first and second electrodes forms the lower exterior, and the inner plate of each of the first and second electrodes forms the lower interior, of two of the lower walls of the cuvette-electrode combination;   wherein a first space is formed within the electroporation cuvette between the inner plates of both the first and second electrodes;   wherein a second space is formed in the electroporation cuvette, above the first space;   wherein the second space is continuous with the first space; and   wherein the structure defining the side boundaries of the second space facilitates the flow of liquids into the first space.   
     
     
         60 . The electroporation cuvette of  claim 59  the volume of the first space may be varied by varying the length of at least one of the interconnecting plates of the first and second electrodes to alter a second fixed distance between the inner and outer plates of at least one of the first and second electrodes. 
     
     
         61 . The electroporation cuvette of  claim 59  wherein the parallel inner and outer plates of the I-beam electrodes have sufficient structural rigidity to retain their shape, meaning that the first fixed distance between the electrodes is maintained during freezing and thawing processes. 
     
     
         62 . The electroporation cuvette of  claim 59  wherein the inner plate of one of the first or second electrodes is configured to function as the electrode surface; and the outer plate of the other of the first or second electrodes is configured to be in contact with one of the lower inside walls of the cuvette. 
     
     
         63 . An electroporation cuvette with an internal structure comprising:
 first and second electrodes positioned within the electroporation cuvette and placed against two of the directly opposing, lower inside walls of the electroporation cuvette, wherein the first and second electrodes are positioned at a first fixed distance apart;
 wherein each of first and second electrodes is in the form of an I-beam, each I-beam comprising (i) inner, and (ii) outer plates that are connected by at least one perpendicular interconnecting plate, the length of which determines the second fixed distance between the inner and outer plates of each of the electrodes; 
   a first space formed between the first and second electrodes; and   a second space formed in the upper portion of the electroporation cuvette, above the first-space;
 wherein the length of at least one of the interconnecting plates of the first and second electrodes may be varied to enable a change in distance between the first and second electrodes, resulting in a change in the volume of the first space; 
 wherein the second space is continuous with the first space; and 
 wherein the structure defining the side boundaries of the second space facilitates the flow of liquids into the first space. 
   
     
     
         64 . A method of using an electroporation cuvette of  claim 44  for transformation of a bacteria or yeast cell, comprising:
 positioning at least one the electrodes inside the cuvette; 
 adjusting and fixing the length of the interconnecting plate; 
 maintaining the fixed length of the interconnecting plate throughout electroporation; 
 wherein the electrode maintains sufficient structural rigidity to control the distance between the electrodes.