User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/10/28: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">PcTF Subcloning in E. coli</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">PcTF Subcloning in E. coli</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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= | =Colony PCR of [http://openwetware.org/wiki/User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli/2013/10/25 Golden Gate Assembly] of BD003 and BD004= | ||
''' | * Pick 10 colonies of each plate and grow in 2 mL LB AMP broth for 6 hours. | ||
* Spin down the cells for 3 minutes in microcentrifuge with top speed. | |||
* Resuspend the pallet in 100μL dH2O. | |||
* Incubate the cells in 98°C for 5 minutes. | |||
* Spin down the cells for 3 minutes in microcentrifuge with top speed. | |||
* Take the supernatent as DNA template for PCR reactions. | |||
* PCR colonies at: 95°C for 3 min. , [95°C 3 min, 45°C 45 sec., 72°C 3 min.] ×35, 72°C 6min. 4°C ∞. | |||
{| {{table}} ; style="text-align:center; width:550px; height:200px;" | |||
|[[Image:20131029 103655 (3).jpg|350px|BD004(HPK promoter) bands shows that only the promoter part is amplified (588bps)]] ||[[Image:20131029 103655 (4).jpg |350px|salam]] | |||
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|BD004('''HPK''' promoter) bands shows that only the promoter part is amplified (588bps) || BD003('''CMV''' promoter) bands shows that only the promoter part is amplified (~ 600bps) | |||
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Latest revision as of 23:31, 26 September 2017
PcTF Subcloning in E. coli | Main project page Previous entry Next entry | ||||
Colony PCR of Golden Gate Assembly of BD003 and BD004
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