User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/04/11

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Current revision (16:57, 11 April 2012) (view source)
 
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BD-112 and BD-113
BD-112 and BD-113
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[[Image:BD-111-Behzad-5-4-12.jpg|thumb|350px|White dashed lines border show the vector backbone and the BD-111 constructed gene(RBS + PcTF).]]
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[[Image:BD112-BD113-Gel.JPG|thumb|350px|2 colonies of BD-112 (1 & 2) and 2 colonies of BD-113 (3 & 4)- White dashed lines border, 1 and 2 show the vector backbone (3426 bp) and the BD-112 constructed gene(T7 + RBS + PcTF + 2 scars) (1196 bp), 3 and 4 show the vector backbone (2079 bp) and the BD-113 constructed gene(T7 + RBS + PcTF + 2 scars) (1196 bp).]]
{| class="wikitable" width=400px  
{| class="wikitable" width=400px  
| Plasmid DNA || 2.0 μl*
| Plasmid DNA || 2.0 μl*
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# Make the (1%) agarose glee and add 15μl  of restricted DNA (PcTF & RBS) in one well and 10 μl  of ladder.
# Make the (1%) agarose glee and add 15μl  of restricted DNA (PcTF & RBS) in one well and 10 μl  of ladder.
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'The size of the constructed gene supposed to be: 1123 + 15 + 6 = 1144 bp'
+
* The size of the constructed gene supposed to be: 1123 + 15 + 6 + 6 + 46 = 1144 bp'

Current revision

PcTF Subcloning in E. coli Main project page
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11/4/12

> Digests (Fermentas FD)
BD-112 and BD-113

2 colonies of BD-112 (1 & 2) and 2 colonies of BD-113 (3 & 4)- White dashed lines border, 1 and 2 show the vector backbone (3426 bp) and the BD-112 constructed gene(T7 + RBS + PcTF + 2 scars) (1196 bp), 3 and 4 show the vector backbone (2079 bp) and the BD-113 constructed gene(T7 + RBS + PcTF + 2 scars) (1196 bp).
2 colonies of BD-112 (1 & 2) and 2 colonies of BD-113 (3 & 4)- White dashed lines border, 1 and 2 show the vector backbone (3426 bp) and the BD-112 constructed gene(T7 + RBS + PcTF + 2 scars) (1196 bp), 3 and 4 show the vector backbone (2079 bp) and the BD-113 constructed gene(T7 + RBS + PcTF + 2 scars) (1196 bp).
Plasmid DNA 2.0 μl*
EcoR1 1.0 μl
Pst1 1.0 μl
10x FastDigest buffer + green loading dye 1.5 μl
dH2O 9.5 μl
  15.0 μl total
Incubate at 37°C for 10 minutes.
  1. Make the (1%) agarose glee and add 15μl of restricted DNA (PcTF & RBS) in one well and 10 μl of ladder.
  • The size of the constructed gene supposed to be: 1123 + 15 + 6 + 6 + 46 = 1144 bp'
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