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Table 4 Comparison of different DNA polymerases for library preparation using LP-PCR-1 and uSGS

From: Ultrasensitive single-genome sequencing: accurate, targeted, next generation sequencing of HIV-1 RNA

Method

Enzyme

cDNA starting copy

Qualified sequences

% Sequences excludeda

Final # sequences

% Population represented

LP-PCR-1

AmpliTaq

Goldb

34,094

11,215

40

6729

20

uSGS

AmpliTaq

Goldb

34,094

11,859

7

11,060

32

LP-PCR-1

Kapa

Uracil+b

34,094

3566

11

3188

9

uSGS

Kapa

Uracil+b

34,094

18,855

2

18,512

54

LP-PCR-1

Platinum

Taqc

14,410

2789

82

558

4

uSGS

Platinum

Taqc

88,560

25,773

10

22,938

26

  1. aFinal number of “super majority” consensus sequences after removal of >2 ambiguous sites likely due to in vitro PCR recombination
  2. bSynthesis of cDNA from WT/Mutant mixture of transcript HIV-1 RNA, divided into 4 parts and parallel libraries sequenced
  3. cSynthesis of cDNA from WT/Mutant mixtures of transcript HIV-1 RNA independently in two separate experiments