RNASep Semi-Prep Column - RPC-99-3015 - Installation, Storage, Regeneration and Precautions

Q: How should the HPLC system be prepared before installing the RNASep™ Semi-Prep column?

Flush the entire system with the intended mobile phase. Pass the mobile phase through a 0.2 µm filter and thoroughly degas it before use.

Q: What are the recommended equilibration and running conditions for RNA loading?

Set the flow rate to 5.0 mL/min, then equilibrate the column with 38% Buffer B for at least 30 minutes before loading the RNA sample. Installation notes also reference maintaining the oven at the targeted temperature.

Q: How do you regenerate and store the column after use?

Regenerate with 100% Solution D after reversing flow and heating to 80 °C (or lower), then flush for 45 minutes, return to normal flow, and continue Solution D until cooled to 50 °C (or lower). For storage, flush with 100% Solution D at 5.0 mL/min for at least 40 minutes, seal using end-plugs, and store in Solution D at room temperature.

Sep 27, 2026

Uploaded by pixelkiddo

RNASep™ Semi-Prep Column RPC-99-3015 provides nonporous PS-DVB (C18-alkylated) packing designed for RNA separation. The guide covers shipment identification, required system flushing with the specified mobile phases, filtration and degassing, recommended UV detection at 260 nm, and maximum column pressure. It details physical specifications, pH range, buffer A/B compositions using TEAA, wash solution D, and step-by-step installation with flow rate and equilibration time. Storage in Solution D, regeneration via reversed flow and temperature cycling, plus key handling and compatibility precautions are included, along with notes on custom buffer manufacture.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages3

Reading Time8 min

AccessFree to download

RNASep™ Semi-Prep Column RPC-99-3015

General requirements

Physical characteristics

Mobile phase and buffers

Installation

Column storage

Column regeneration procedure

Column precautions

Custom buffer manufacture

RNASep™ Semi-Prep Column Column Care Information Catalog Number: RPC-99-3015 The ADS Biotec RNASep™ Semi-Prep Column contains nonporous polystyrene-divinylbenzene (PS-DVB) copolymer beads that are approximately 2µm in size and are alkylated with C-18 chains. The polymer packing is designed for the separation of RNA. The column should be installed and used in accordance with the following instructions. GENERAL Each column is shipped with an attached label identifying column type, serial number, and flow direction. Be sure this tag is kept on the column in case further details are needed about this specific column. Before installing the column, the entire system should be flushed with the mobile phase to be used. The mobile phase should be passed through a 0.2􀁐m filter and thoroughly degassed. Maximum pressure on this column should not exceed 4000 psi. The typical detection system uses UV absorbance @ 260nm. PHYSICAL CHARACTERISTICS Dimensions: 30 x 150 mm Packing Material: Alkylated PS-DVB beads, C18 Ionic Form: NA pH Range: 0-14 Column Volume ~ 106 mL Column Void Volume ~ 67 mL MOBILE PHASE The RNASep Semi-Prep column was designed for optimal separation using ADS Biotec WAVE Optimized® Buffer system. TEAA Buffer A- (PN: 553421) 0. 1M Triethylammonium Acetate (TEAA) pH 7.0 TEAA Buffer B- (PN: 553422) 0. 1M TEAA, 25% Acetonitrile pH 7.0 Column Wash Solution D- (PN: 553423) 75% Acetonitrile INSTALLATION -Before installing the column, thoroughly flush the HPLC system with 100 % Solution D (for 30 minutes @ 2.5 mL/min) . Best results will be obtained with the flow direction as indicated by the arrow on the column. -Securely attach the column and turn on the column oven to targeted oven temperature. -Set Flow Rate to 5.0 mL/min. -Equilibrate the column with 38% Buffer B for at least 30 min before loading RNA sample. COLUMN STORAGE Be sure to flush RNASep™ Semi-Prep column with 100% Solution D @ 5.0 mL/min, (~ 1.5 column volume) for at least 40 minutes before removing column for storage. The column must be stored in Solution D. Prior to storage, seal the column using the column end-plugs before storing at room temperature. COLUMN REGENERATION PROCEDURE To prolong the life of the RNASep™ Semi-Prep Column it may be necessary to regenerate using Solution D. 1. Reverse the flow direction of the column. 2. Set oven temperature to 80 °C (or lower than 80 °C as column heater allows) . 3. Flush column with 100% Solution D @ 3.0 mL/min for 45 minutes. 4. Set oven temperature to 50 °C. 5. Reverse the flow direction of the column-back to normal direction. NOTE: Column will be HOT. 6. To prepare column for storage, continue with Solution D until column has cooled to 50 °C (or lower) before removing. 7. To prepare column for a new sample, equilibrate well using running buffers e.g. 50% Buffer A, 50% Buffer B @ 2.5 mL/min for at least 30 minutes at the targeted oven temperature. COLUMN PRECAUTIONS No warranty exists for this specialized column. Please note the following precautions in using the RNASep™ Semi-Prep column: • Use only HPLC grade acetonitrile, <0 .005 AU (UV absorbance) at 260 nm. ADS Biotec Buffers and\or acetonitrile are recommended. • Use only HPLC grade water that has a resistivity of at least 18 MΩ purity with < 15 ppb T.O.C. (total organic carbon) and must not be autoclaved. • Avoid using the following materials with the column: • Bovine Serum Albumin • Autoclaved Water • Mineral Oil • Formamide • Proteinase K • High Molecular weight stabilizers such as polyethylene glycol (1% max) • Detergents such as Triton X100, NP40, Tween 20 and SDS/SLS (1% max) • Glycerol (2% max) • DMSO (10% max) • Betaine (1 .25-2.5M max) • Reagents containing metals such as iron, chromium, or nickel Custom Buffer Manufacture Contact us for your custom needs for mobile phase manufacturing of ready-to-use buffer solutions to be produced within our proven ISO 1

FAQs

How should the HPLC system be prepared before installing the RNASep™ Semi-Prep column?

Flush the entire system with the intended mobile phase. Pass the mobile phase through a 0.2 µm filter and thoroughly degas it before use.

What are the recommended equilibration and running conditions for RNA loading?

Set the flow rate to 5.0 mL/min, then equilibrate the column with 38% Buffer B for at least 30 minutes before loading the RNA sample. Installation notes also reference maintaining the oven at the targeted temperature.

How do you regenerate and store the column after use?

Regenerate with 100% Solution D after reversing flow and heating to 80 °C (or lower), then flush for 45 minutes, return to normal flow, and continue Solution D until cooled to 50 °C (or lower). For storage, flush with 100% Solution D at 5.0 mL/min for at least 40 minutes, seal using end-plugs, and store in Solution D at room temperature.

Sep 27, 2026

Uploaded by pixelkiddo