Nucleic acid library preparation using electrophoresis

A method of preparing a library of tagged nucleic acid fragments from a target double-stranded nucleic acid, the method comprising:

• (a) first providing an electrophoresis system comprising:
• a single electrophoresis gel matrix having a first end and a second end, a width, and a length between the first and second ends, wherein all reactants other than a sample to be reacted are present in the electrophoresis gel matrix prior to electrophoresis operation and reactants other than the sample are not added or removed from the electrophoresis gel matrix during a sample run prior to elution;
• a first portion proximate to the first end and containing a first transposase cofactor for a transposase, wherein the first transposase cofactor is a metallic ion;
• a second portion proximate to the second end and containing a second transposase cofactor for the transposase, the second transposase cofactor having an electrical charge opposite to that of the first transposase cofactor, wherein the second transposase cofactor is a polynucleotide transposase adaptor comprising a double-stranded transposon end sequence and a single-stranded tag region;
• a reaction portion between the first portion and the second portion and containing the transposase;
• and
• a pair of positive and negative electrodes arranged at the first and second ends, respectively;
• (b) after step (a), applying the sample to the reaction portion, wherein the sample comprises a target double-stranded nucleic acid;
• (c) after step (b), applying a first electric field between the pair of electrodes to drive the first transposase cofactor from the first portion into the reaction portion and drive the second transposase cofactor from the second portion into the reaction portion to form an activated transposome complex comprising the transposase, the first transposase cofactor, and the second transposase cofactor;
• and
• (d) after step (c), incubating the target double-stranded nucleic acid with the activated transposome complex in the reaction portion under conditions sufficient to fragment the target double-stranded nucleic acid and tag the nucleic acid fragments, thereby generating a library of tagged nucleic acid fragments.

A method of preparing a library of tagged nucleic acid fragments from a target double-stranded nucleic acid, the method comprising:

• (a) first providing an electrophoresis system comprising:
• a single electrophoresis gel matrix having a first end and a second end, a width, and a length between the first and second ends, wherein all reactants other than a sample to be reacted are present in the electrophoresis gel matrix prior to electrophoresis operation and reactants other than the sample are not added or removed from the electrophoresis gel matrix during a sample run prior to elution such that multiple reactions take place within the electrophoresis gel matrix;
• a first portion containing a first transposase cofactor for a transposase, wherein the first transposase cofactor is a metallic ion;
• a second portion containing a second transposase cofactor for the transposase, the second transposase cofactor having an electrical charge opposite to that of the first transposase cofactor, wherein the second transposase cofactor is a polynucleotide transposase adaptor comprising a double-stranded transposon end sequence and a single-stranded tag region;
• a third portion containing a lysis reagent;
• a fourth portion containing the transposase;
• a reaction portion between the first portion and the second portion;
• and
• a first pair of positive and negative electrodes arranged at the first and second ends, respectively;
• and
• a second pair of positive and negative electrodes arranged at opposite sides of the width at one of the first end or the second end;
• (b) after step (a), applying the sample to the reaction portion, wherein the sample comprises whole cells containing a target double-stranded nucleic acid;
• (c) after step (b), applying a first electric field to drive the lysis agent into and/or through the reaction portion to lyse the cells, thereby applying the target double-stranded nucleic acid to the reaction portion;
• (d) after step (b) applying a second electric field to drive the first transposase cofactor from the first portion into the reaction portion, drive the second transposase cofactor from the second portion into the reaction portion, and/or drive the transposase from the transposase portion to the reaction portion to form an activated transposome complex comprising the transposase, the first transposase cofactor, and the second transposase cofactor, wherein the first electric field is also applied during at least a portion of step (d);
• and
• (e) after step (d), incubating the target double-stranded nucleic acid with the activated transposome complex in the reaction portion under conditions sufficient to fragment the target double-stranded nucleic acid and tag the nucleic acid fragments, thereby generating a library of tagged nucleic acid fragments.

An electrophoresis device comprising:

• a single electrophoresis gel matrix having a first end and a second end, a width, and a length between the first and second ends, wherein all reactants other than a sample to be reacted are present in the electrophoresis gel matrix prior to electrophoresis operation and reactants other than the sample are not added or removed from the electrophoresis gel matrix during a sample run prior to elution;
• a first portion proximate to the first end and containing a first transposase cofactor, wherein the first transposase cofactor is a metallic ion;
• a reaction portion distant to the first end and containing a transposome complex comprising a transposase and a polynucleotide transposase adaptor comprising a double-stranded transposon end sequence and a single-stranded tag region;
• and
• a pair of positive and negative electrodes arranged at the first and second ends, respectively.