Skip to main content
Figure 1 | Biological Procedures Online

Figure 1

From: Simultaneous splicing of multiple DNA fragments in one PCR reaction

Figure 1

Principles of our simultaneous splicing overlap extension PCR (SSOE-PCR). Step 1, two fragments with overlap sequences are amplified by normal PCR after choosing better outmost primers (Forward primer and Reverse primer) and overlap primers. Step 2, a program of five cycles is employed at higher annealing temperature after correctly mixing DNA polymerase buffer, dNTPs, few of the two fragments, outermost primers and Pfu DNA polymerase as well as adding water to certain volume. Some fused DNAs (DNA1 plus DNA2) will be synthetized. Step 3, a continuous program of 30 cycles following step 2 is edited and running at lower annealing temperature through normal PCR protocols.

Back to article page