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Solid Phase PCR (SP-PCR) (CAT#: STEM-MB-0219-WXH)

Introduction

Solid-phase PCR (SP-PCR) is a unique PCR technique that allows amplification of target nucleic acids on a solid support where one or both primers are immobilized on the surface.<br />The spatial separation of the primers minimizes significantly undesirable primer interactions, thereby preventing the formation of primer-dimers and allowing higher multiplexing amplification.




Principle

This method uses PCR primers modified with biotea at the 5' end to bring biotin to the ends of the amplified DNA sequence, followed by hybridization of the amplified sequence with an isotope-labeled probe in solution. DNA fragment complementary to the amplified DNA fragment provides the detection signal.

Applications

• Specific detection of pathogen DNA
• DNA sequencing
• Extraction (purification) of specific nucleic acid fragments
• Solid phase targeted mutagenesis

Procedure

General procedure:
1. Denaturation
2. Annealing
3. Extension