Method for nucleic acid amplification

A composition comprising:

• at least one target nucleic acid molecule and
• an amplicon library,
• wherein the amplicon library comprises a plurality of polynucleotides obtained from amplification of the at least one target nucleic acid molecule,
• wherein at least some of the polynucleotides comprise a terminator nucleotide,
• wherein no more than 50% of a cumulative fraction of polynucleotides comprises sequences of at least 80% of a cumulative fraction of sequences of the at least one target nucleic acid molecule.

A method of sequencing a target nucleic acid molecule, the method comprising:

• a. contacting a sample comprising the target nucleic acid molecule, at least one amplification primer, at least one nucleic acid polymerase, and a mixture of nucleotides, wherein the mixture of nucleotides comprises at least one terminator nucleotide which terminates nucleic acid replication by the polymerase, and
• b. amplifying the target nucleic acid molecule to generate a plurality of terminated amplification products, wherein the replication proceeds by strand displacement replication;
• c. removing at least one terminator nucleotide from the terminated amplification products;
• d. ligating the molecules obtained in step (c) to adaptors, thereby generating a library of amplification products;
• and
• e. sequencing the library of amplification products.

A method of sequencing a single cell, the method comprising:

• a. providing a cell lysate from the single cell;
• b. contacting the cell lysate with at least one amplification primer, at least one nucleic acid polymerase, and a mixture of nucleotides, wherein the mixture of nucleotides comprises at least one terminator nucleotide which terminates nucleic acid replication by the polymerase, and
• c. amplifying the target nucleic acid molecule to generate a plurality of terminated amplification products, wherein the replication proceeds by strand displacement replication;
• d. removing at least one terminator nucleotide from the terminated amplification products;
• e. ligating the molecules obtained in step (d) to adaptors, thereby generating a library of amplification products;
• and
• f. sequencing the library of amplification products.

A method of determining the mutagenicity of an environmental condition, the method comprising:

• a. exposing cells to the environmental condition;
• b. isolating single cells from the population;
• c. providing a cell lysate from a single cell;
• d. contacting the cell lysate with at least one amplification primer, at least one nucleic acid polymerase, and a mixture of nucleotides, wherein the mixture of nucleotides comprises at least one terminator nucleotide which terminates nucleic acid replication by the polymerase, and
• e. amplifying the target nucleic acid molecule to generate a plurality of terminated amplification products, wherein the replication proceeds by strand displacement replication;
• f. removing at least one terminator nucleotide from the terminated amplification products;
• g. ligating the molecules obtained in step (f) to adaptors, thereby generating a library of amplification products;
• and
• h. sequencing the library of amplification products, and
• i. comparing the sequences of amplification products to at least one reference sequence to identify mutations.