User:Jose Fabricio Lopez/Notebook/Logbook/2011/08/01: Difference between revisions
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=Objectives= | =Objectives= | ||
* Digestion of[[ User:Jose Fabricio Lopez/Notebook/Logbook/2011/07/29 | Plasmid extraction]] pSB1AK3/BBa_B0015 for Standard Assembly. | * Digestion of[[ User:Jose Fabricio Lopez/Notebook/Logbook/2011/07/29 | Plasmid extraction]] pSB1AK3/BBa_B0015 for Standard Assembly. | ||
* PCR for | * PCR for BBa_P1003 with preffix and suffix primers. | ||
* See in gel digestion and product of PCR. | * See in gel digestion and product of PCR. | ||
* Clean today´s PCR product. | * Clean today´s PCR product. | ||
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Enzymes inactivation= 20 min 65°C | Enzymes inactivation= 20 min 65°C | ||
==PCR for | ==PCR for BBa_P1003== | ||
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| 6 | | 6 | ||
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|PCR product( | |PCR product(BBa_P1003) | ||
| 40 | | 40 | ||
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Revision as of 02:11, 5 August 2011
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Objectives
Digestion
PCR for BBa_P1003
Denaturation step: 30seg 94°C Annealing step: 30seg 55°C Extension/elongation step: 40seg 72°C Final elongation 30seg 72°C Final hold: 3hrs 4°C Gel of PCR and digestion
PCR product purificationRoche´s kit. Final dilution= 60μL Gel of PCR purified Vs Plasmid CutThis gel just show concentration between backbone and insert and concentration for Double Digestion of insert(Purified PCR product) Double Digestion of PCR productDouble Digestion with EcoRI-HF and SpeI
StatusSuccess!!!
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