Synthesis of a primer test sequence using StratoSpheres DNA Application Note Sl-02001

About this manual

The document details the preparation of oligonucleotides for PCR applications. The synthesis was performed on an Applied Biosystems 392 DNA/RNA Synthesizer using standard chemistry, followed by cleavage with ammonium hydroxide and deprotection at 55 °C overnight.

Following synthesis, HPLC analysis was conducted to evaluate the performance of the StratoSpheres DNA dT support. The analysis utilized a Pellicular SAX column with a specific gradient of Tris-HCl and Ammonium Chloride, detecting results at UV 260 nm.

What's inside

• Introduction
• Oligonucleotide Synthesis
• HPLC Analysis

Frequently asked questions

What is the specific test sequence used to evaluate the StratoSpheres DNA support?

The document uses a 20-mer test sequence with the sequence 5' ATA CCG ATT AAG CGA AGT TT 3'.

What temperature and duration are required for deprotecting the side chain protecting groups?

Deprotection is accomplished by heating to 55 °C overnight.

What is the flow rate used during the HPLC analysis of the oligonucleotides?

The specified flow rate for the analysis is 1.5 mL/min.