Abstract: In one embodiment of the invention, a method is provided for preparing plasmid from host cells which contain the plasmid, comprising: (a) providing a plasmid solution comprised of unligatable open circular plasmid; (b) reacting the unligatable open circular plasmid with one or more enzymes and appropriate nucleotide cofactor(s), such that unligatable open circular plasmid is converted to 3?-hydroxyl, 5?-phosphate nicked plasmid; (c) reacting the 3?-hydroxyl, 5?-phosphate nicked plasmid with a DNA ligase and DNA ligase nucleotide cofactor, such that 3?-hydroxyl, 5?-phosphate nicked plasmid is converted to relaxed covalently closed circular plasmid; and (d) reacting the relaxed covalently closed circular plasmid with a DNA gyrase and DNA gyrase nucleotide cofactor, such that relaxed covalently closed circular plasmid is converted to negatively supercoiled plasmid. In other embodiments, DNA gyrase is replaced with reverse DNA gyrase or reaction (d) is not performed.

Abstract: In one embodiment of the invention, a method is provided for preparing plasmid from host cells which contain the plasmid, comprising: (a) providing a plasmid solution comprised of unligatable open circular plasmid; (b) reacting the unligatable open circular plasmid with one or more enzymes and appropriate nucleotide cofactors, such that unligatable open circular plasmid is converted to 3?-hydroxyl, 5?-phosphate nicked plasmid; (c) reacting the 3?-hydroxyl, 5?-phosphate nicked plasmid with a DNA ligase and DNA ligase nucleotide cofactor, such that 3?-hydroxyl, 5?-phosphate nicked plasmid is converted to relaxed covalently closed circular plasmid; and (d) reacting the relaxed covalently closed circular plasmid with a DNA gyrase and DNA gyrase nucleotide cofactor, such that relaxed covalently closed circular plasmid is converted to negatively supercoiled plasmid. In other embodiments, DNA gyrase is replaced with reverse DNA gyrase or reaction (d) is not performed.

Abstract: In one embodiment of the invention, a method is provided for preparing plasmid from host cells which contain the plasmid, comprising: (a) providing a plasmid solution comprised of unligatable open circular plasmid; (b) reacting the unligatable open circular plasmid with one or more enzymes and appropriate nucleotide cofactors, such that unligatable open circular plasmid is converted to 3?-hydroxyl, 5?-phosphate nicked plasmid; (c) reacting the 3?-hydroxyl, 5?-phosphate nicked plasmid with a DNA ligase and DNA ligase nucleotide cofactor, such that 3?-hydroxyl, 5?-phosphate nicked plasmid is converted to relaxed covalently closed circular plasmid; and (d) reacting the relaxed covalently closed circular plasmid with a DNA gyrase and DNA gyrase nucleotide cofactor, such that relaxed covalently closed circular plasmid is converted to negatively supercoiled plasmid. In other embodiments, DNA gyrase is replaced by reverse DNA gyrase or reaction (d) is not performed.

Abstract: In one embodiment of the invention, a method is provided for preparing plasmid from host cells which contain the plasmid, comprising: (a) providing a plasmid solution comprised of unligatable open circular plasmid; (b) reacting the unligatable open circular plasmid with one or more enzymes and appropriate nucleotide cofactors, such that unligatable open circular plasmid is converted to 3?-hydroxyl, 5?-phosphate nicked plasmid; (c) reacting the 3?-hydroxyl, 5?-phosphate nicked plasmid with a DNA ligase and DNA ligase nucleotide cofactor, such that 3?-hydroxyl, 5?-phosphate nicked plasmid is converted to relaxed covalently closed circular plasmid; and (d) reacting the relaxed covalently closed circular plasmid with a DNA gyrase and DNA gyrase nucleotide cofactor, such that relaxed covalently closed circular plasmid is converted to negatively supercoiled plasmid. In other embodiments, DNA gyrase is replaced by reverse DNA gyrase or reaction (d) is not performed.

Abstract: A universal sequential logic circuit is constructed from a rectilinear array of elementary logic "cells", with a relatively large number of logic states embodied in a relatively small array. The set of states from a state-machine description of the logic function desired to be performed is compiled into a software association of cellular array states with each state-machine state (4), and the set of transitions from the state-machine description is compiled into a software association of logical connections between cells. The cellular array performs the state-machine function under software control. The rectilinear array (1a) generally embodies one bit of cellular array state information for each row of logic cells, stored in diagonal cells of the array called "memory cells" (2). Non-diagonal cells of the array called "function cells" (3) are controlled by stored software, which controls the transfer of cellular array state information from each row to each other.

Abstract: A programmable logic device includes means for operating a computing element to compile a set of state-machine states in an incompletely specified state-machine. The state-machine states are compiled into a set of cellular array states in a rectilinear format of columns and rows. Multiple memory cells are located on a main diagonal. Function cells are located removed from the main diagonal for transferring information between the memory cells. Compatible sets of sequences are formed of sequences which have non-equal effect. Compatible sets are processed to form a closed cover. A distinct memory cell is then assigned to each compatible set constituting that closed cover. The closed cover can be formed selectively by having a compatible set consist of either a single entry of one sequence, two or more sequences, being at least a pair of sequences, a maximal compatible set or less than maximal compatible set from a pair of compatibles.

Abstract: A method for sequencing nucleic acid polymers is provided in which the polymer to be sequenced acts as a template for the production of a complementary polymer by a polymerase enzyme. The template polymer is introduced into a polymerization environment in which production of the complementary polymer will occur if appropriate nucleotides are provided. The nucleotides are then provided to the polymerization environment one at a time in individual feedstocks. If the nucleotide in a feedstock is complementary to the next base in the template polymer, i.e., the unpaired base closest to the growing end of the complementary polymer, polymerization will occur lengthening the complementary polymer and releasing PPi. By separately recovering each feedstock and analyzing it for the presence of PPi, the sequence of the complementary polymer and thus the template polymer is determined.