User:Karmella Haynes/Notebook/BioBrick cloning/2015/04/22: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Karmella's BioBrick Cloning</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Karmella's BioBrick Cloning</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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* Submit to DNASU for sequencing | * Submit to DNASU for sequencing | ||
** Order Number is: 10068 | ** Order Number is: 10068 (4/23/15) | ||
# g53-2, f (P139) | # g53-2, f (P139) - Confirmed | ||
# g53-2, r (P139) | # g53-2, r (P139) - low Phred 20 score | ||
# g54, f (P139) | # g54, f (P139) - Confirmed | ||
# g54, r (P139) | # g54, r (P139) - low Phred 20 score | ||
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| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| rowspan="7" | <u>Expected:</u><br>1,2. AubR/MRV = ~3200, 886<br>3,4. BjaR/MRV = ~3200, 776<br>5,6. BraR/MRV = ~3200, 776<br>7,8. RpaR/MRV = ~3200, 779<br>MRV (empty) = ~3200 | | rowspan="7" | <u>Expected:</u><br>1,2. AubR/MRV = ~3200, 886<br>3,4. BjaR/MRV = ~3200, 776<br>5,6. BraR/MRV = ~3200, 776<br>7,8. RpaR/MRV = ~3200, 779<br>MRV (empty) = ~3200 | ||
| rowspan="7" | | | rowspan="7" | [[Image:KAH042315_gel1.jpg|250px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
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| DNA(plasmid) || 3.0 μL | | DNA(plasmid) || 3.0 μL | ||
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| EcoRI || 1.0 | | EcoRI || 1.0 | ||
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| | | XbaI || 1.0 | ||
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| dH<sub>2</sub>O || 8.5 | | dH<sub>2</sub>O || 8.5 | ||
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Conclusions | Conclusions | ||
* | * AubR/MRV - success! Use clone 1 (lane 1) | ||
* BjaR/MRV - success! Use clone 1 (lane 3) | |||
* BraR/MRV - failed. Pick more clones | |||
* RpaR/MRV - success! Use clone 2 (lane 8) | |||
Latest revision as of 00:55, 27 September 2017
Karmella's BioBrick Cloning | Main project page Previous entry Next entry | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
04/22/15
Rene - minipreps & sequencing
LCR - assembly attempt #1
LCR Reactions (PCR fragments / oligo bridges)
Thermocycler: BioRad dual block
Ryan - Receiver cloning confirmations
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