Team:UNITN-Trento/Notebook/Labposts/06/44

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{
{
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"date" : "2013-06-24",
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"date" : "2013-06-20",
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"author" : "caterina-michele",
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"author" : "emil-gabriele",
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"title" : " Gel, gel, gel..of BBa_K1065101",
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"title" : "Linearization of psB1C3(Failed) and screening of SAM sinthethase",
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"content" : "<html>After the incula that Michele made on Sunday of araCpBAD with BBa_K1065100 (BMST1 in pSB1C3) we did the screening of the minipreps.Before obtaining a good result we tried three different gels.As you can see from these pictures (METTI IMMAGINI) the first two gels made from the same digestion were not lucky while the last, performed with different agarose concentration (1.5% instead of 1%) gave a nice result!</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Screening BBa_K1065101|<html><center><img src=\"https://static.igem.org/mediawiki/2013/7/7a/Tn-20130624-BMST1_ARAcpbad_PSB1C3.jpg\" height=\"300px\" /></center></html>}}",
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"content" : "<html>We quantified yesterday inocula (1:1, 1:2, 1:4 of SAMsynthetase and R0010, both in pSB1C3).</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Quantification results|<html><center><table class=\"tn-sp-table\"><tr><th>Sample</th><th>Quantification (ng/&micro;l)</th></tr><tr><td>1:1 A</td><td>293.0</td></tr><tr><td>1:1 B</td><td>135.7</td></tr><tr><td>1:1 C</td><td>178.0</td></tr><tr><td>1:1 D</td><td>139.3</td></tr><tr><td>1:1 E</td><td>262.0</td></tr><tr><td>1:1 F</td><td>147.5</td></tr><tr><td>1:1 G</td><td>212.5</td></tr><tr><td>1:1 H</td><td>210.2</td></tr><tr><td>1:2 A</td><td>141.0</td></tr><tr><td>1:2 B</td><td>123.6</td></tr><tr><td>1:4 A</td><td>207.8</td></tr><tr><td>1:4 B</td><td>127.2</td></tr><tr><td>R0010 A</td><td>131.7</td></tr><tr><td>R0010 B</td><td>119.6</td></tr><tr><td>R0010 C</td><td>371.0</td></tr><tr><td>R0010 D</td><td>307.9</td></tr><tr><td>R0010 E</td><td>313.8</td></tr></table></center></html>}}<html>Then, we screened the products of ligation (1:1, 1:2, 1:4 and R0010): digestion with EcoRI-HF and PstI-HF following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">digestion protocol</a> and agarose gel 1%.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|1% Agarose Gel Image|<html><center><table class=\"tn-sp-table\"><tr><th>Loading Scheme</th></tr><tr><td><i>Empty</i></td></tr><tr><td>1kb ladder</td></tr><tr><td><i>Empty</i></td></tr><tr><td>1:1 D</td></tr><tr><td>1:1 E</td></tr><tr><td><i>Empty</i></td></tr><tr><td>1:2 B</td></tr><tr><td><i>Empty</i></td></tr><tr><td>1:4 A</td></tr><tr><td><i>Empty</i></td></tr><tr><td>R0010 E</td></tr></table><img src=\"https://static.igem.org/mediawiki/2013/5/54/Tn-20130620-ET_GG_screening_SAM_R0010_Failed.jpg\" width=\"450px\" /> <!-- Sams primo screening--></center></html>}}{{:Team:UNITN-Trento/Templates/Styles/Spoiler|1% Agarose Gel Image|<html><center><table class=\"tn-sp-table\"><tr><th>Loading scheme</th></tr><tr><td>1kb ladder</td></tr><tr><td>R0010 A</td></tr><tr><td>R0010 B</td></tr><tr><td>R0010 C</td></tr><tr><td>R0010 D</td></tr><tr><td>R0010 E</td></tr></table><img src=\"https://static.igem.org/mediawiki/2013/1/1d/Tn-20130620-ET_screening_psB1C3SAM_FAILED.jpg\" width=\"450px\" /> <!-- Sams 1:1 and PCR(last 3) --></center></html>}}{{:Team:UNITN-Trento/Templates/Styles/Spoiler|1% Agarose Gel Image|<html><center><table class=\"tn-sp-table\"><tr><th>Loading scheme</th></tr><tr><td>R0010 B</td></tr><tr><td>R0010 C</td></tr><tr><td>R0010 D</td></tr><tr><td>1kb ladder</td></tr><tr><td>1:4 A</td></tr><tr><td>1:4 B</td></tr><tr><td>1:2 A</td></tr><tr><td>1:2 B</td></tr></table><img src=\"https://static.igem.org/mediawiki/2013/a/aa/Tn-20130620-GG_R0010_pSB1C3SAM_FAILED.jpg\" width=\"450px\" /> <!-- R0010 --></center></html>}}{{:Team:UNITN-Trento/Templates/Styles/Spoiler|1.5% Agarose Gel Image|<html><center><table class=\"tn-sp-table\"><tr><th>Loading scheme</th></tr><tr><td>1kb ladder</td></tr><tr><td>R0010 A</td></tr><tr><td>R0010 B</td></tr><tr><td>R0010 C</td></tr><tr><td>R0010 D</td></tr><tr><td>R0010 E</td></tr></table><img src=\"https://static.igem.org/mediawiki/2013/8/8b/Tn-20130620-GG_R0010_screening_FAILED.jpg\" width=\"450px\" /> <!-- R0010 bis --></center></html>}}<html>In these images we can only see the band of the backbone and no insert!</html>",
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"tags" : "BBa_K1065101"
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"tags" : "SAMsynthetase"
}
}

Latest revision as of 07:52, 3 October 2013

{ "date" : "2013-06-20", "author" : "emil-gabriele", "title" : "Linearization of psB1C3(Failed) and screening of SAM sinthethase", "content" : "We quantified yesterday inocula (1:1, 1:2, 1:4 of SAMsynthetase and R0010, both in pSB1C3).

Quantification results
SampleQuantification (ng/µl)
1:1 A293.0
1:1 B135.7
1:1 C178.0
1:1 D139.3
1:1 E262.0
1:1 F147.5
1:1 G212.5
1:1 H210.2
1:2 A141.0
1:2 B123.6
1:4 A207.8
1:4 B127.2
R0010 A131.7
R0010 B119.6
R0010 C371.0
R0010 D307.9
R0010 E313.8
Then, we screened the products of ligation (1:1, 1:2, 1:4 and R0010): digestion with EcoRI-HF and PstI-HF following the digestion protocol and agarose gel 1%.
1% Agarose Gel Image
Loading Scheme
Empty
1kb ladder
Empty
1:1 D
1:1 E
Empty
1:2 B
Empty
1:4 A
Empty
R0010 E
1% Agarose Gel Image
Loading scheme
1kb ladder
R0010 A
R0010 B
R0010 C
R0010 D
R0010 E
1% Agarose Gel Image
Loading scheme
R0010 B
R0010 C
R0010 D
1kb ladder
1:4 A
1:4 B
1:2 A
1:2 B
1.5% Agarose Gel Image
Loading scheme
1kb ladder
R0010 A
R0010 B
R0010 C
R0010 D
R0010 E
In these images we can only see the band of the backbone and no insert!", "tags" : "SAMsynthetase" }

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