Haynes Lab:Notebook/Engineering PC-TFs/2013/01/29: Difference between revisions
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==Summary== | ==Summary== | ||
| <br><font size=3>'''Digestion/ ligation reaction'''</font> | | <br><font size=3>'''Digestion/ ligation reaction'''</font> | ||
* <br>'''Dilute the purified PCR product to 20 fmol/μL''' | |||
* Measure ng/μL of the purified sample. | * Measure ng/μL of the purified sample. | ||
* The volume of purified DNA (x) you will need to dilute in a final volume of 20 μL = 20 μL final volume * 20 fmols/μL * '''length in bp''' * 650 fg/fmol ÷ 1,000,000 fg/ng ÷ '''measured ng/μL''' | * The volume of purified DNA (x) you will need to dilute in a final volume of 20 μL = 20 μL final volume * 20 fmols/μL * '''length in bp''' * 650 fg/fmol ÷ 1,000,000 fg/ng ÷ '''measured ng/μL''' | ||
** Formula: x = '''length in bp''' ÷ '''measured ng/μL''' * 0.26 | ** Formula: x = '''length in bp''' ÷ '''measured ng/μL''' * 0.26 | ||
* '''Perform BsmBI/ T4 ligase mediated assembly''' | |||
* BsmBI cuts the DNA fragments and creates complementary overhangs. | * BsmBI cuts the DNA fragments and creates complementary overhangs. | ||
* Complementary sticky ends anneal via base pairing. | * Complementary sticky ends anneal via base pairing. |
Revision as of 00:03, 1 February 2013
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Summary |
Digestion/ ligation reaction
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