DNA,RNA

DNA fragments digested by Hae III were analyzed using IEC DEAE-420N (a column for weak anion exchange chromatography packed with non-porous gel).
DEAE-420N is an old type of DEAE3N-4T.

Sample : phi-X-174-RF DNA fragment

No.
base pairs
MW (x 105daltons)
1.
1353
8.79
2.
1078
7.01
3.
872
5.67
4.
603
3.92
5.
310
2.02
6B.
281
1.83
6A.
271
1.76
7.
234
1.52
8.
194
1.26
9.
118
0.77
10.
72
0.47

 

Column       : Shodex IEC DEAE-420N (4.6mmID*35mm)
Eluent       : (A); 25mM Phosphate buffer(pH5.2) 
               (B); (A) + 1M NaCl
               Linear gradient: 0min to 20min, 75% (B) to 100% (B) 
Flow rate    : 1.0mL/min
Detector     : UV(260nm)
Column temp. : 30deg-C

IEC DEAE-825 packed with porous gel can allow larger sample loads than IEC DEAE3N-4T packed with non-porous gel.

Sample :
Transfer ribonucleic acid, tRNA (from E.Coli)

Column       : Shodex IEC DEAE-825 (8.0mmID*75mm)
Eluent       : (A); 20mM Sodium phosphate buffer(pH6.6) + 250mM KCl + 5M Urea + 0.5mM EDTA 
               (B); (A) + 250mM KCl
               Linear gradient: 0min to 150min, (A) to (B)
Flow rate    : 1.0mL/min
Detector     : UV(260nm) 
Column temp. : Room temp.

Transfer ribonucleic acids were analyzed using OHpak SB-804 HQ (a column for GFC separation).
Please refer to Transfer RNA (3) (SB-805 HQ)

Sample :
1. 23s-RNA(E.Coli MRE 600)
2. 16s-RNA(E.Coli MRE 600)
3. 5s-RNA(E.Coli MRE 600)
4. Transfer ribonucleic acid, tRNA(E.Coli MRE 600)

Columns      : Shodex OHpak SB-804 HQ (8.0mmID*300mm) x 2
Eluent       : 0.1M CH3COONa buffer(pH7.0) + 0.3M NaCl + 1mM EDTA
Flow rate    : 0.1mL/min
Detector     : UV(260nm)
Column temp. : Room temp.

Transfer ribonucleic acids were analyzed using OHpak SB-805 HQ (a column for GFC separation).
Refer to Transfer RNA (2) (SB-804 HQ)

Sample :
1. 23s-RNA(E.Coli MRE 600)
2. 16s-RNA(E.Coli MRE 600)
3. 5s-RNA(E.Coli MRE 600)
4. Transfer ribonucleic acid, tRNA(E.Coli MRE 600)

Columns      : Shodex OHpak SB-805 HQ  (8.0mmID*300mm) x 2
Eluent       : 0.1M CH3COONa buffer(pH7.0) + 0.3M NaCl + 1mM EDTA
Flow rate    : 0.1mL/min
Detector     : UV(260nm)
Column temp. : Room temp.

Transfer ribonucleic acids were analyzed using IEC DEAE-420N (a column for weak anion exchange chromatography packed with non-porous gel).
DEAE-420N is an old type of DEAE3N-4T.

Sample : tRNA (E.Coli MRE600)

Column       : Shodex IEC DEAE-420N (4.6mmID*35mm)
Eluent       : (A); 25mM Tris-HCl(pH7.5) (B); (A) + 1M NaCl
               Linear gradient: 20%(B) to 50%(B), 0 to 20min
Flow rate    : 0.5mL/min
Detector     : UV(260nm)
Column temp. : 60deg-C

Ribosomel RNA is analyzed using IEC DEAE-420N (a column for weak anion exchange chromatography packed with non-porous gel).
DEAE-420N is an old type of DEAE3N-4T.

Sample : 5s-RNA (E.Coli MRE600)

Column       : Shodex IEC DEAE-420N (4.6mmID*35mm)
Eluent       : (A); 25mM Tris-HCl buffer(pH7.5) 
               (B); (A) + 1M NaCl
               Linear gradient: 0min to 15min, 10% (B) to 100% (B)
Flow rate    : 1.5mL/min
Detector     : UV(260nm)
Column temp  : 60deg-C

Ribosomel RNA were analyzed using IEC DEAE-420N (a column for weak anion exchange chromatography packed with non-porous gel).
DEAE-420N is an old type of DEAE3N-4T.

Sample : 16s+23s-RNA (E.Coli MRE600)

Column       : Shodex IEC DEAE-420N (4.6mmID*35mm)
Eluent       : (A); 25mM Tris-HCl buffer(pH7.5) 
               (B); (A) + 1M NaCl
               Linear gradient: 0min to 60min, 25% (B) to 100% (B)
Flow rate    : 0.5mL/min
Detector     : UV(260nm)
Column temp. : 30deg-C

Ribosomal RNA was analyzed using Asahipak GS-620 7G (a column for GFC separation). For general HPLC analysis, a high flow rate of 0.5-1.0mL/min or over is generally used. Whereas, for the GFC analysis of high molecular weight components, it is recommended that the analysis be made at a flow rate as low as possible, e.g. 0.1mL/min to prevent molecular chains from being cut off and to permit a high resolution.
GS-620 7G is an old type of GS-620 HQ.

Sample :
1. Ribosomal RNA-23S
2. Ribosomal RNA-16S
3. Ribosomal RNA-5S

Column       : Shodex Asahipak GS-620 7G (7.5mmID*500mm)
Eluent       : 0.1M Sodium phosphate buffer(pH7.0) + 0.3M NaCl
Detector     : UV(260nm)
Column temp. : 30deg-C

Plasmid DNA fragments were analyzed using Asahipak GS-710 7G (a column for GFC separation).
GS-710 7G is an old type of GF-710 HQ.

Sample : Plasmid DNA Fragment

Columns      : Shodex Asahipak GS-710 7G (7.5mmID*500mm) x 2
Eluent       : 50mM Tris-HCl buffer(pH7.2) + 300mM NaCl + 2mM EDTA-2Na
Flow rate    : 0.1mL/min
Detector     : UV(260nm)
Column temp. : 30deg-C

Crude gamma phage DNA fragments were analyzed using Asahipak GS-510 7G (a column for GFC separation).
GS-510 7G is an old type of GF-510 HQ.

Sample : Crude gamma phage DNA fragment

Column     : Shodex Asahipak GS-510 7G (7.5mmID*500mm)
Eluent     : 0.1M Tris-HCl buffer(pH7.5) + 0.2M NaCl
Flow rate  : 0.6mL/min
Detector   : UV(260nm)

J. Hirabayashi and K. Kasai, Anal. Biochem., 178 (1989) 336.

Aqueous SEC column, OHpak SB-806 HQ was used for the analysis of polyadenylic acid.

Sample : 20micro-L
Polyadenylic acid, poly[A]n 0.1%
(Wako Pure Chemical Industries, Ltd.
code:101011 POLY-ADENYLIC ACID)

Column       : Shodex OHpak SB-806 HQ (8.0mmID*300mm)
Eluent       : 10mM Sodium phosphate buffer(pH4.6) 
Flow rate    : 1.0mL/min
Detector     : UV(260nm)
Column temp. : 30deg-C