Methods of cDNA preparation
The present disclosure provides methods and systems for amplifying and analyzing nucleic acid samples. The present disclosure provides methods for preparing cDNA and/or DNA molecules and cDNA and/or DNA libraries using modified reverse transcriptases.
1. A method for preparing a complementary deoxyribonucleic acid (cDNA) molecule comprising:
(a) partitioning a cell and a non-naturally occurring reverse transcriptase having at least 95% identity to SEQ ID NO: 20 in a partition, which cell comprises ribonucleic acid (RNA) molecules;
(b) releasing said RNA molecules from said cell in said partition; and
(c) in said partition, using said non-naturally occurring reverse transcriptase to synthesize a complementary deoxyribonucleic acid (cDNA) library from said RNA molecule, which non-naturally occurring transcriptase synthesizes said cDNA library at a processivity of 20 nucleotides or longer.
2. The method of claim 1 , wherein said non-naturally occurring reverse transcriptase has at least 100% identity to SEQ ID NO: 20.
3. The method of claim 1 , wherein said partition further comprises one or more acceptor nucleic acid molecules.
4. The method of claim 1 , further comprising performing said method on a heterogeneous plurality of template nucleic acid molecules comprising a plurality of distinct ribonucleic acid (RNA) molecules.