81
rpc
RPC
HIGHLIGHTS
Porous silica with 25 nm (250 A) pore size
C18 bonded phase in TSKgel OligoDNA RP suited for
Oligonucleotides
C1 bonded phase in TSKgel TMS-250 suited for proteins
TSKgel TMS-250 is exhaustively and repeatedly reacted with trimethyl
silyl groups. Standard nomenclature designates the bonded phase as
C1. This wide-pore column is recommended for the analysis of proteins.
TSKgel OligoDNA RP contains a monomeric C18 bonded phase that
is not endcapped and has a relatively low carbon content of 10%.
It is ideal for the purification and analysis of oligonucleotides (up to
500-mer), RNAs, and DNA fragments. It possesses high-resolving
power for octamers of similar sequence.
Applications
TSKgel TMS-250
TSKgel TMS-250 is ideal for the separation of proteins which
exhibit sharp peaks relative to wide-pore C8 or C18 columns. It can
accommodate even large proteins, such as aldolase (158 kDa). The
resolution of proteins on TSKgel TMS-250 columns is shown in Figure 3.
TSKgel OligoDNA RP
The semi-preparative isolation of a 49-mer oligonucleotide from the
crude synthetic reaction mixture using a 7.8 mm ID TSKgel OligoDNA-RP
column is shown in Figure 4. The purity of the isolated oligonucleotide
was subsequently verified on an analytical 4.6 mm ID TSKgel OligoDNA-
RP column.
RP Columns for Biomolecules TSKgel OligoDNA RP / TMS-250
figure 3
High resolution protein separation on TSKgel TMS-250
Retention time (minutes)
0
30
15
2
1
3
4
7
5
6
Column:
TSKgel TMS-250, 4.6mm ID x 7.5cm
Mobile phase: 60min linear gradient from 20% to 95%
CH
3
CN in 0.05% TFA, pH2.2
Flow rate:
0.61mL/min
Detector response (AU)
Column: TSKgel TMS-250, 4.6 mm ID x 7.5 cm L;
Sample: 5 μg each of: 1. ribonuclease A, 2. cytochrome C, 3. lysozyme,
4. bovine serum albumin, 5. aldolase, 6. carbonic anhydrase, 7. ovalbumin;
Elution: 60 min (TMS-250) linear gradient from 20% to 95% CH
3
CN in 0.05%
TFA, pH 2.2; Flow rate: 0.61 mL/min; Detection: UV @ 220 nm
ORDERING INFORMATION
Part #
Description
ID
Length
Particle
Number
Flow rate
Maximum
(mm)
(cm)
size (µm)
theoretical
(mL/min)
pressure
plates
range
drop (MPa)
TSKgel stainless steel columns
0013352 OligoDNA RP, 25 nm
4.6
15.0
5
7,000 0.6 - 1.0
12.0
0013353 OligoDNA RP, 25 nm
7.8
15.0
5
7,000 2.0 - 3.0
12.0
0007190 TMS-250, 25 nm
4.6
7.5
10
1,500 0.5 - 0.8
2.0
figure 4
High resolution protein separation on TSKgel TMS-250
Retention time (minutes)
0
10
20
0
10
20
30
A. Isolation
B. Purity check
Detector response (AU)
Detector response (AU)
Columns: A. TSKgel OligoDNA-RP, 5 µm, 7.8 mm ID × 15 cm, B. TSKgel
OligoDNA-RP, 5 µm, 4.6 mm ID × 15 cm
Mobile phase: A. 120 min linear gradient from 6.25% to 25% CH
3
CN (7.8 mm
ID) column; B. 90 min linear gradient from 7.5% to 25% CH
3
CN (4.6 mm ID)
column, both in 0.1 mol/L ammonium acetate, pH 7.0,
Flow rate: A. 2.8 mL/min (7.8 mm ID) B. 1.0 mL/min (4.6 mm ID),
Detection: UV @ 260 m, Sample: synthetic 49-mer oligonucleotide,
d(AGCTTGGGCTGCAGGTCGTCTCTAGAGGATCCCCGGGCGAGCTCGAATT)
