Agilent TapeStation

All RNA and DNA quality control for library construction is done on an Agilent TapeStation 4150. This service is only offered as part of quantification for library preparation and sequencing and is not offered as a stand-alone service.

RNA

High Sensitivity RNA ScreenTape AssayRNA ScreenTape Assay
sample volume required2 μL1 μL
sizing range25 - 6000 bp25 - 6000 bp
sensitivity0.1 ng/μL5 ng/μL
quantitative precision15% CV10% CV
quantitative accuracy±30%±20%
quantitative range0.5 - 10 ng/μL25 - 500 ng/μL
RINe functional range1 - 25 ng/μL25 - 500 ng/μL
maximum buffer concentration in sample10 mM Tris, 1 mM EDTA200 mM Tris, 20 mM EDTA, or 50 mM NaCl

DNA

High Sensitivity DNA 1000 ScreenTape AssayDNA 1000 ScreenTape Assay
sample volume required2 μL1 μL
sizing range35 - 1000 bp35 - 1000 bp
typical resolution35 - 300 bp: 15%, 300 - 1000 bp: 10%)35 - 300 bp: 15%, 300 - 1000 bp: 10%
sensitivity0.005 ng/μL0.1 ng/μL
sizing precision5% CV5% CV
sizing accuracy±10% (w/ electronic ladder: ±20%)±10% (w/ electronic ladder: ±20%)
quantitative precision15% CV0.1 - 1 ng/μL: 15% CV, 1 - 50 ng/μL: 10% CV
quantitative accuracy±20%±20%
quantitative range0.01 - 1 ng/μL0.1 - 50 ng/μL
maximum buffer concentration in sample7 mM KCl, 20 mM phosphate buffer,
20 mM guanidine-HCl, 80 mM NaCl,
20 mM C2H4O2-Na
60 mM phosphate buffer, 20 mM KCl,
60 mM guanidine-HCl, 240 mM NaCl,
60 mM C2H4O2-Na

Invitrogen Qubit

Qubit assays utilize fluorescent molecules that are usually highly selective for a particular nucleic acid (e.g. dsDNA over RNA, RNA over DNA) and accurate in the presence of DNA, salts, solvents, proteins, and free nucleotides. Prior to library construction, input RNA and DNA samples are quantified using the Qubit 3 Fluorometer. This service is only offered as part of quantification for library preparation and sequencing and is not offered as a stand-alone service.

RNA

RNA High Sensitivity AssayRNA Broad Range Assay
sample volume required1-20 μL1-20 μL
sample conc. range0.25 - 100 ng/μL1 - 1000 ng/μL
sample amount range4 - 200 ng10 - 1200 ng
assay conc. range (total)0.02 - 1 ng/μL0.05 - 6 ng/μL
assay conc. range (highest confidence)0.025 - 0.50 ng/μL0.10 - 5 ng/μL
maximum contaminant conc. in assay10 mM NaCl, 2 mM MgCl2,
10 mM C2H4O2-Na/C2H4O2-H3N
1% EtOH, 0.1% Phenol,
1X NTPs/oligos/ssDNA/dsDNA
...
10 mM NaCl, 2 mM MgCl2,
10 mM C2H4O2-Na/C2H4O2-H3N
0.1% EtOH, 0.1% Phenol,
1X NTPs/oligos/ssDNA/dsDNA
...

DNA

1X dsDNA High Sensitivity AssaydsDNA Broad Range Assay
sample volume required1-20 μL1-20 μL
sample conc. range0.01 - 100 ng/μL0.10 - 1000 ng/μL
sample amount range0.10 - 120 ng2 - 2000 ng
assay conc. range (total)0.0005 - 0.60 ng/μL0.01 - 10 ng/μL
assay conc. range (highest confidence)0.001 - 0.50 ng/μL0.01 - 5 ng/μL
maximum contaminant conc. in assay50 mM NaCl, 1 mM MgCl2,
30 mM C2H4O2-Na, 25 mM C2H4O2-H3N
1% EtOH, 0.1% Phenol,
1X dNTPs/ssDNA/RNA
...
10 mM NaCl, 2 mM MgCl2,
10 mM C2H4O2-Na/C2H4O2-H3N
1% EtOH, 0.1% Phenol,
100 μM dNTPs, 6X RNA, 1X ssDNA, 3X oligos
...

LightCycler 96

For some types of libraries, quantification prior to sequencing is done on the Roche LightCycler 96 to ensure equimolar pooling and appropriate loading concentrations.

Library Quantification by qPCR

Library quantification kits provide accurate and reliable absolute quantification of libraries prepared for sequencing on Illumina platforms. The kit functions across a wide range of library types, concentrations, fragment distributions and GC content.