Template:SBB-Protocols PCR3
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SOEing PCR
- Set up PCR reactions according to your construction file as normal 33uL reactions as described in Cloning by PCR
- For each PCR, load 6uL of PCR product premixed with 4uL of loading buffer in a single well of a 1% agarose gel
- Cut out the bands, put them into a single 1.5mL microcentrifuge tube
- Add 650uL of ADB Buffer
- Proceed with the Zymo Gel Purification procedure
- Elute the DNA in 50uL of water
- Set up your second round of PCR as a normal 33uL reaction using the eluted mixture of fragments as template
