Table 1

Comparison of current sequencing library quantification methods

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


* Limit of quantification for ssDNA 500-mer;

** Manufacturer does not specify this value as LOD or LOQ

White et al. BMC Genomics 2009 10:116   doi:10.1186/1471-2164-10-116

Open Data