User:Jose Fabricio Lopez/Notebook/Logbook/2011/08/01: Difference between revisions
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Roche´s kit. | Roche´s kit. | ||
Final dilution= 60μL | Final dilution= 60μL | ||
==Gel of PCR purified Vs Plasmid Cut== | |||
This gel just show concentration between backbone and insert and concentration for Double Digestion of insert(Purified PCR product) | |||
[[Image:PCRpurified vs plasmidDigested.1August.2011.JPG|120px]] | |||
==Double Digestion of PCR product== | |||
Double Digestion with EcoRI-HF and SpeI | |||
{|border=1 | |||
!Reactives | |||
!μL | |||
|-- | |||
|Buffer4 10x | |||
| 6 | |||
|-- | |||
|BSA 10x | |||
| 6 | |||
|-- | |||
|PCR product(Prom+Kana) | |||
| 40 | |||
|-- | |||
|SpeI | |||
| 1 | |||
|-- | |||
|EcoRI-HF | |||
| 1 | |||
|-- | |||
| H<sub>2</sub>O | |||
| 6 | |||
|-- | |||
| Total | |||
| 60 | |||
|} | |||
==== | ==== |
Revision as of 00:43, 2 August 2011
iGEM 2011 | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Objectives
Digestion
PCR for Kanamycin
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
==Status: Success!!!
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