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Table 1 Comparison of current sequencing library quantification methods

From: Digital PCR provides sensitive and absolute calibration for high throughput sequencing

Method:

Nanodrop

Capillary GE

Ribogreen

Real-time PCR

UT-qPCR

UT-digital qPCR

Detection Chemistry:

UV absorption

Intercalating fluorophore

Intercalating fluorophore

Syber Green I Intercalating fluorophore

Hydrolysis probe (Taqman)

Hydrolysis probe (Taqman)

Companies:

Thermo Scientific

Aligent, Bio-Rad

Invitrogen

Many

Many

Fluidigm

LOQ:*

2 ng**

(7.2 billion copies)

25 ng

(91 billion copies)

1 ng

(3.6 billion copies)

(0.3 fg)

1000 copies

(0.03 fg)

100 copies

(0.03 fg)

100 copies

Quantification Modality:

Mass/absolute

Mass/relative

Mass/relative

Mass/relative

Molecules/relative

Molecules/absolute

Quantification Standard:

No standard necessary

Required – calibrated by mass

Required – calibrated by mass

Required – calibrated by mass

Required – calibrated by mass

No standard necessary

Reference:

nanodrop.com

Ricicova, M. et al (2003)

Jones, Lj et al. (1998)

Simpson (2000); Meyer (2008)

Zhang (2003); This work

Kalinina (1997); This work

  1. * Limit of quantification for ssDNA 500-mer;
  2. ** Manufacturer does not specify this value as LOD or LOQ