Pathlength Correction Factors; Direct Uv Assays For Dna, Rna, Oligonucleotides And Protein Concentration; Concentration - Thermo Scientific Evolution 200 Series User Manual

18
nanoCell

Pathlength Correction Factors

Direct UV Assays for DNA, RNA, Oligonucleotides and Protein Concentration

Calculation Factor
Assay Unit
1.0 cm Pathlength
dsDNA ng/μL
ssDNA ng/μL
RNA ng/μL
Oligos ng/μL
Protein 280 nm mg/mL
Protein 205 nm mg/mL
Direct UV Assays for DNA, RNA, Oligonucleotides and Protein Concentration
Assay Unit Equation
260/280 μg/mL DNA and Protein Concentrations determined using:
DNA conc = [(A
DNA/
Protein conc = [(A
Protein
260:280 Ratio =
Where:
A = absorption at 260 nm
260
A = absorption at 280 nm
280
A = background absorption at 320 nm
ref
260/230 μg/mL DNA and Protein Concentrations determined using:
DNA conc = [(A
DNA/
Protein conc = [(A
Protein
(Phenol)
260:230 Ratio =
Where:
A = absorption at 260 nm
260
A = absorption at 230 nm
230
A = background absorption at 320 nm
ref
Warburg- mg/mL Protein concentration is determined using:
Christian Protein conc = [(A
Where:
A = absorption at 280 nm
280
A = absorption at 260 nm
260
288 Evolution 200 Series User Guide
Calculation Factor
1.0 mm Pathlength
nanoCell
50 500
37 370
40 400
33 330
1 10
31 310
- A )f - (A - A )f ]
260 ref 1 230 ref 2
- A )f - (A - A )f
230 ref 3 260 ref 4
A – A
260 ref
--------------------------- -
A – A
280 ref
- A )f - (A - A )f ]
260 ref 1 230 ref 2
- A )f - (A - A )f
230 ref 3 260 ref 4
A – A
260 ref
--------------------------- -
A – A
230 ref
x f ) - (A x f )]
280 1 260 2
Calculation Factor
0.2 mm Pathlength Reference
nanoCell
2500 Maniatis, T; Fritch, E. F.; Sambrook, J.,
Molecular Cloning: A Laboratory Handbook,
3650
2nd Edition, Cold Spring Harbor Laboratory
Press, Plainview, NY 1989, Vol. 3.
2000
1650
50 Fasman, D. G., Ed. Practical Handbook of
Biochemistry and Molecular Biology, CRC
1550
Press, Boston 1992.
Stoscheck, C. M., Methods Enzymol, 1990,
182, 50-69.
Calculation
Factors Factor
1.0 mm 1.0 mm
Pathlength Pathlength
nanoCell
f = 62.9 f = 629
1 1
f = 36.0 f = 360
2 2
] f = 1,552 f = 15,520
3 3
f = 757.3 f = 7,573
4 4
f = 49.1 f = 491
1 1
f = 3.48 f = 34.8
2 2
] f = 183 f = 1,830
3 3
f = 75.8 f = 758
4 4
1.55 15.5
0.76 7.6
Calculation
Factor
0.2 mm References
Pathlength
nanoCell
f = 3,145 Warburg, O.;
1
f = 1,800 Christian, W.
2
f = 77,600 Biochem Z. 1942,
3
f = 37,865 310, 384-421.
4
Groves, W. E.;
Davis, F. C., Jr.;
Sells, B. H., Anal.
Biochem. 1968, 22,
195-210.
f = 12,300 Kalb, V. F., Jr.;
1
f = 870 Bernlohr, R.W.,
2
f = 45,750 Anal. Biochem.
3
f = 18,950 1977, 82, 362-371.
4
3,875
Warburg, O.;
1,900
Christian, W.,
Biochem Z. 1942,
310, 384-421.
Thermo Scientific