Template:SBB-Protocols PCR3: Difference between revisions
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===SOEing PCR=== | ===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 8uL of PCR product premixed with 1uL 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 | |||
Revision as of 21:44, 3 February 2009
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 8uL of PCR product premixed with 1uL 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
