Method and kit for preparing complementary dna

A kit for preparing complementary deoxyribonucleic acid (cDNA) comprising:

• a cDNA synthesis primer configured to hybridize to a ribonucleic acid (RNA) molecule to enable synthesis of a cDNA strand complementary to at least a portion of the RNA molecule to form an RNA-cDNA intermediate;
• and
• a template switching oligonucleotide (TSO) comprising an amplification primer site, an identification tag, a unique molecular identifier (UMI) and multiple predefined nucleotides, wherein the TSO is configured to act as a template in a template switching reaction comprising extension of the cDNA strand to form an extended cDNA strand complementary to the at least a portion of the RNA molecule and the TSO.

The method according to any of the preceding claims, wherein the fragmenting comprises tagmenting to produce tagged fragments.

The method according to any of the preceding claims, wherein the method further comprises sequencing the first population of 5′ UMI comprising fragments and a second population of internal fragments.

A kit for preparing nucleic acid fragments, the kit comprising:

• a cDNA synthesis primer configured to hybridize to a ribonucleic acid (RNA) molecule to enable synthesis of a cDNA strand complementary to at least a portion of the RNA molecule to form an RNA-cDNA intermediate and comprising a reverse amplification primer site;
• and
• a template switching oligonucleotide (TSO) comprising an amplification primer site, an identification tag, a unique molecular identifier (UMI) and multiple predefined nucleotides, wherein the TSO is configured to act as a template in a template switching reaction comprising extension of the cDNA strand to form an extended cDNA strand complementary to the at least a portion of the RNA molecule and the TSO.