Berkeley LBL/JoyceNotebook
From 2007.igem.org
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4. [[Berkeley_LBL/Digestion|Digestion for PCR Products]] using specific restriction enzymes | 4. [[Berkeley_LBL/Digestion|Digestion for PCR Products]] using specific restriction enzymes | ||
| + | {| border = | ||
| + | |- | ||
| + | !''bchD'' | ||
| + | |NdeI, BamHI | ||
| + | |||
| + | |- | ||
| + | !''bchI'' | ||
| + | |NdeI, BglI | ||
| + | |} | ||
Revision as of 19:37, 25 October 2007
Contruction of pET3a Derivatives Containing bchD and bchI
1. Amplify Rhodobacter's gene and introduce restriction sites by PCR (Using Phusion Polymerase), which allow cloning PCR fragments into pET3a
| bchD | bchI | |
|---|---|---|
| length of gene fragment | 1695 b.p. | 1005 b.p. |
| Restriction sites introduced when amplified by PCR | NdeI, BamHI | NdeI, BglI |
|
|
2. Do Analytic Digestion to confirm that the length of the PCR fragments are the same as the corresponding target gene fragments.
3. Remove any leftover PCR enzyme in the samples by PCR Clean Up/Purification.
4. Digestion for PCR Products using specific restriction enzymes
| bchD | NdeI, BamHI |
|---|---|
| bchI | NdeI, BglI |
