Abstract:
An electrophoretic separating and blotting apparatus, which includes a pressure block, two glass plates of different heights, a partition frame, a rack, a positioning unit, a cell, a blotting cassette, and two electrode plates. The component parts are alternatively used for electrophoretic gelation as well as blotting, enabling electrophoretic gelation process and blotting process to be efficiently and economically proceeded within a short length of time.

Description:
BACKGROUND OF THE INVENTION 
     The present invention relates to an electrophoretic separating and blotting apparatus, and more particularly to such an apparatus, which is practical for use in electrophoretic gelation process, as well as blotting process. 
     Electrophoresis is a technique commonly used to separate protein and DNA. According to conventional methods, a gel is cast between two plates (in vertical) or on a flat bed (in horizontal), and inserted in between buffer/electrode chambers in an electrophoretic apparatus. A buffer solution is added to the buffer/electrode chambers, and a small amount of sample solution is loaded in wells in the gel. Glycerin and “tracing” dye are put in the wells or added to the sample solution. An electric current is then given to the gel. After tracing dye has been run through the length of the gel, the electric current is stopped, and the gel is removed from the apparatus for analysis to measure the transfer of the molecules of the sample. The gel can be colored by, for example, a dye, enabling the dye to be bonded to protein or DNA. The gel can then be dried, preserved, or photographed, enabling the distribution of the molecules of the sample to be recorded for permanent visual check. After electrophoresis, the gel must be taken out of the electrophoretic apparatus, and then delivered to a blotting apparatus for blotting. Because the electrophoretic apparatus and the blotting apparatus are two independent apparatus, they must be separately prepared. Because the electrophoretic procedure and the blotting procedure must be performed separately in the electrophoretic apparatus and the blotting apparatus, it is complicated and time-consuming to achieve electrophoretic and blotting operations. 
     SUMMARY OF THE INVENTION 
     The present invention has been accomplished to provide an electrophoretic separating and blotting apparatus, which eliminates the aforesaid drawbacks. It is the main object of the present invention to provide an electrophoretic separating and blotting apparatus, which can be arranged into different modes for running an electrophoretic process as well as a blotting procedure economically and efficiently. 
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS 
     FIGS. 1,  2 ,  2 A and  3  are exploded and sectional assembly views of a part of an electrophoretic separating and blotting apparatus according to the present invention. 
     FIGS. 4 and 4A are elevational and sectional side views of a part of the present invention, showing the apparatus arranged for casting. 
     FIGS. 5,  6  and  6 A are elevational, exploded and sectional side views of the present invention for the steps of loading and running. 
     FIGS. 7,  8 ,  9 ,  10 ,  10 A and  11  are elevational, exploded and sectional side views of the present invention, showing the apparatus arranged for blotting. 
    
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT 
     An electrophoretic gel separation method is a technique commonly used to separate protein and DNA. Charge carried molecules are movable under the guide of an electric field. By means of the porous nature of a gel and under the driving of an electric field, molecules are separated or arranged subject to the order of molecular size. 
     Electrophoresis includes the steps of gelation (alignment, casting), sample-loading, and running. The performance of one step affects the success of a next step. 
     Referring to FIGS. from  1  through  6 , an electrophoretic separating an blotting apparatus in accordance with the present invention comprises a pressure block  1 , two glass plates  11  and  11 ′, two spacers  12 , a comb  13 , a partition frame  2 , a rack  3 , a positioning unit  4 , a cell  5 , a transfer membrane  7 , two pieces of filter paper  8 , a blotting cassette  9 , and two plate electrodes  10  and  10 ′. 
     The glass plates  11  and  11 ′ have different heights. The spacers  12  and  12 ′ are bilaterally arranged between the glass plates  11  and  11 ′. The comb  13  is arranged in the difference of elevation between the glass plates  11  and  11 ′. 
     The partition frame  2  comprises a first side jamb  21 , a second side jamb  21 ′ arranged in parallel to the first side jamb  21 , a bottom rail  22  connected between the first side jamb  21  and the second side jamb  21 ′ at the bottom, a transverse plate  25  connected between the side jambs  21  and  21 ′ and spaced above the bottom rail  22 , two positioning rods  26  and  26 ′ respectively outwardly raised from the side jambs  21  and  21 ′ at the top for fastening to two opposite lateral sides of the cell  5 . The side jambs  21  and  21 ′ each have a positioning groove  23  longitudinally disposed at an outer side, and respectively coupled to two vertical sidetracks  31  inside the rack  3 . The bottom rail  22  has a positioning groove  221  longitudinally disposed at a bottom side in communication with the positioning groove  23  at each side jamb  21  and  21 ′. The second side jamb  21 ′ has a vertical through hole  211 ′ through which a negative electrode is inserted with a conductor and fastened to a locating block  24  between the bottom rail  22  and the second side jamb  21 ′, forming a negative electric zone. The first side jamb  21  has a vertical through hole  211  through which a positive electrode is inserted with a conductor and extended to the positioning groove  23  adjacent to the bottom rail  22 , forming a positive electric zone. A protruding index  27  is provided at the positioning rod  26 ′ at the second side jamb  21 ′ for guiding the installation of the cover  52  of the cell  5  into position to prevent an electric connection error. Two locating grooves  28  are provided at the front and back sides of the partition frame  2 , and extended through the side jambs  21  and  21 ′ and the bottom rail  22 . A flexible gasket  29  is respectively fastened to each locating groove  28 . Each locating groove  28  has two sloping ends  281  extended upwardly outwards toward the top at each side jamb  21  and  21 ′. After installation of the gaskets  29  in the locating grooves  28 , the two distal ends of each gasket  29  are respectively forced outwards and firmly stopped at the glass plates  11  and  11 ′. 
     The rack  3  comprises two vertical tracks  31  bilaterally disposed on the inside for the positioning of the side jambs  21  and  21 ′, a plurality of holding down elements  33  adjusted to evenly press on the pressure block  1  against the glass plates  11  and  11 ′, symmetrical positioning portions  34  provided at two opposite peripheral side walls  32  thereof at a bottom side for positioning on the positioning unit  4 , a plurality of protruding bearing portions  35  respectively formed integral with the positioning portions  34 , a plurality of vertical locating grooves  36  bilaterally provided on the inside for the positioning of other members used during blotting, two receptacle portions  37  bilaterally disposed on the outside, two terminal holder plates  38  respectively inserted in the receptacle portions  37 , and two banana plugs  39  respectively installed in the terminal holder plates  38 . The terminal holder plates  38  each comprise two positioning grooves  381  disposed at two opposite lateral sides and respectively coupled to one receptacle portion  37 , a positioning rod  382  and a bearing rod  383  horizontally extended from the top side thereof in reversed directions, and a protruding indicator  384  for recognition of polarity. The banana plugs  39  are respectively mounted on the bearing rod  383  at each terminal holder plate  38 . 
     The positioning unit  4  comprises a recessed portion  42  at the top side wall thereof on the middle, a carrier plate  43  mounted in the recessed portion  42 , a gasket  44  for positioning in the recessed portion  42  during an electrophoretic gelation operation, a plurality of locating holes  41  spaced around the recessed portion  42  for receiving the positioning portions  34  of the rack  3 , two barrels  45  formed integral with the top side wall and transversely disposed at two opposite sides of the recessed portion  42 , and two pivot members  47  respectively coupled to the barrels  45 . The coupling blocks  45  each have a notch  46 . The pivot members  47  each comprise a pivot shaft  471  respectively and axially inserted into the barrels  45 , two cams  473  fixedly provided at two distal ends of the pivot shaft  471 , and a finger rod  472  perpendicularly disposed on the middle and respectively extended out of the notch  46  at each barrel  45 . The cams  473  each have a protruding portion  474 . 
     The cell  5  comprises a casing  51  for holding the aforesaid parts and a buffer, and a cover  52  covered on the casing  51 . The casing  51  comprises two coupling holes  511  at two opposite lateral side walls thereof. The cover  52  comprises a top handle  521 , two cylindrical terminal receptacles  522  and  522 ′ raised from the top side wall thereof at two opposite sides of the top handle  521 , and a bottom constraint block  523  raised from the bottom side wall thereof for correct polarity positioning of the cover  52  on the casing  51 . 
     The blotting cassette  9  is comprised of two symmetrical perforated plates, namely, the first perforated plate  91  and the second perforated plate  92 , and a sliding lock  93 . The first perforated plate  91  comprises a plurality of foot elements  911  symmetrically raised from a back side wall thereof for supporting on a table top, a plurality of plug strips  912  downwardly extended from the bottom side thereof, and a track  913  forwardly raised from the top side thereof. The second perforated plate  92  comprises a plurality of different sizes of adjustment plug holes  921  provided at the bottom side thereof for receiving the plug strips  912  of the first perforated plate  91 , a top rail  922 , and a notch  923  at the top rail  922  on the middle for receiving the track  913  of the first perforated plate  91 . The plug strips  912  are selectively inserted to the adjustment plugholes  912  subject to the desired pitch between the perforated plates  91  and  92 . After insertion of the plug strips  912  into the adjustment plugholes  912 , the track  913  is positioned in the notch  923 , and the sliding lock  93  is coupled to the top rail  922  to secure the track  913  to the top rail  922  (see FIG.  7 ). 
     The electrode plates  10  and  10 ′ each comprise a plurality of wire grooves  102  arranged in parallel, and a plurality of through holes  101  respectively disposed at two distal ends of each wire groove  102 . Electrode conductors  103  are respectively inserted through the through holes  101 , and arranged in the wire grooves  102  (see FIG.  9 ). 
     Referring to FIGS. 1,  2  and  2 A, before electrophoretic gelation, all the parts are installed, and fixedly secured in place. During alignment, the carrier plate  43  is mounted in the recessed portion  42  at the positioning unit  4 , then the partition frame  2  is mounted in the rack  3 , and then the rack  3  with the partition frame  2  are mounted on the positioning unit  4  by plugging the positioning portions  34  into the respective locating holes  41 , and then the pressure block  1  is inserted into the rack  3 , and then the spacers  12  and  12 ′ are positioned in between the two glass plates  11  and  11 ′ and inserted with the glass plates  11  and  11 ′ into the rack  3  between the partition frame  2  and the pressure block  1 , and then the holding down elements  33  are fastened up to press the pressure block  1  against the glass plates  11  and  11 ′, enabling the glass plates  11  and  11 ′ to be positively supported on the gasket  29 , and then the rack  3  is removed from the positioning unit  4 . After removal of the rack  3  from the positioning unit  4 , the carrier plate  43  is taken away (see FIGS. 3,  4  and  4 A), then the gasket  44  is put in the recessed portion  42 , and then the rack  3  is installed in the positioning unit  4  again, enabling the partition frame  2  and the inner area of the rack  3  to be allocated at the gasket  44 , and then the finger rods  472  of the pivot members  47  are respectively turned inwards to force the protruding portion  474  of each cam  473  against the bearing portions  35  of the rack  3 . At this time, a gelatin fluid is filled into the space defined between the glass plates  11  and  11 ′ to produce a first layer of gel, and then the comb  13  is installed in the difference of elevation between the glass plates  11  and  11 ′. After gelation of the gelatin fluid, a second layer of gel is made, After gelation of the second layer of gel, the comb  13  is taken away, and a plurality of sample wells  61  are formed on the gel  6  at the top side (see FIG.  7 ). Thereafter, the rack  3  is removed from the positioning unit  4  and put in the cell  5 , enabling the mounting rods  26  and  26 ′ of the partition frame  2  to be respectively engaged into the coupling holes  511  at the casing  51  of the cell  5 , and then a buffer is filled in the casing  51  and between two gels (or partition frames  2 ), causing the negative electrode, the gels  6 , the buffer and the positive electrode form a loop. Thereafter, a sample is loaded to the wells  61  at the gel  6 , then the cover  52  is covered on the casing  51  (see FIGS. 5,  6  and  6 A), and then the banana plugs  39  are electrically connected, causing the sample to be arranged in order in the holes in the gel  6 . After running for a certain length of time, the partition frame  2  and the rack  3  are removed from the casing  51 , then the holding down elements of the rack  3  are loosened, and then the pressure block  1  and the glass plates  11  and  11 ′ are removed from the rack  3 , and then the gel  6  is taken away from the rack  3 , and then the partition frame  2  is separated from the rack  3 , thus the blotting operation can then be proceeded. 
     Referring to FIGS. from  7  through  11 , before starting the blotting operation, the two pieces of filter paper  8  and the transfer membrane  7  are pre-treated to prevent the occurrence of bubbles in the filter paper  8 , the transfer membrane  7  or the gel during blotting, then the transfer membrane  7  is adhered to one side of the gel  6 , and then the two pieces of filter paper  8  are respectively adhered to the other side of the gel  6  and the transfer membrane  7 , and then the gel  6  and the transfer membrane  7  with the filter paper  8  are fastened to the blotting cassette  9 . Thereafter, the terminal holder plates  38  are respectively fastened to the insertion slots  37  at the rack  3 , then the rack  3  is put in the casing  51  of the cell  5 , enabling the mounting rods  382  of the terminal holder plates  38  to be respectively fastened to the coupling holes  511  at the casing  51 , and then the blotting cassette  9  is put in the rack  3 , and then the electrode plates  10  and  10 ′ are respectively inserted into the vertical locating grooves  36  in the rack  3 , and the banana plugs  39  are respectively connected to the electrode plates  10  and  10 ′, and then the buffer is poured in the cell  5 , and then the cover  52  is covered on the casing  51 , and the blotting process is started after connection to power supply. After blotting, the rack  3  is removed from the cell  5 , then the blotting cassette  9  is dismantled, and the blotted transfer membrane is obtained. 
     While only one embodiment of the present invention has been shown and described, it will be understood that various modifications and changes could be made thereto without departing from the spirit and scope of the invention.