User:Karmella Haynes/Notebook/BioBrick cloning/2015/05/21: 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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| colspan="2"| | | colspan="2"| | ||
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<!-- Precede finished items with a checkmark ✓ --> | <!-- Precede finished items with a checkmark ✓ --> | ||
* Cas-tone - Assemblies, H3's + MV11 | * Cas-tone - Assemblies, H3's + MV11 | ||
* Ryan - pick pBra-BraR-MRV colonies | |||
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|- bgcolor=#cfcfcf | |- bgcolor=#cfcfcf | ||
| Sample || OD260 || 260/280 || ng/μL | | Sample || OD260 || 260/280 || ng/μL | ||
|- | |||
| 1. MV11 (S/N) || 0.03 || 1.879 || 13.5 | |||
|- | |- | ||
| 2. H3-1-40 PCR || 0.03 || 1.879 || 29.8 | | 2. H3-1-40 PCR || 0.03 || 1.879 || 29.8 | ||
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| 4. H3-1-80 PCR || 0.047 || 1.865 || 46.8 | | 4. H3-1-80 PCR || 0.047 || 1.865 || 46.8 | ||
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| | | 5. H3-1-116 PCR || 0.06 || 1.887 || 59.57 | ||
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| | | 6. H3-1-136 PCR || 0.062 || 1.932 || 62.0 | ||
|} | |} | ||
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* Ligations | * Ligations | ||
** > 2:1 | ** > 2:1 ratio calculations... | ||
** 433 bp largest insert / 9506 bp vector * 2 * | ** 433 bp largest insert / 9506 bp vector * 2 * 50 ng vector = 4.5 ng | ||
** 129 bp smallest insert / 9506 bp vector * 2 * | ** 129 bp smallest insert / 9506 bp vector * 2 * 50 ng vector = 1.4 ng | ||
{| {{table}} cellspacing="3" <!-- Ligation rxn table --> | {| {{table}} cellspacing="3" <!-- Ligation rxn table --> | ||
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| Insert DNA || 0.5 || --- | | Insert DNA || 0.5 || --- | ||
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| Vector DNA || | | Vector DNA || 3.5 || 3.5 | ||
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| 2x lgn buf (Roche) || | | 2x lgn buf (Roche) || 5.0 || 5.0 | ||
|- | |- | ||
| T4 ligase (NEB) || 1.0 || 1.0 | | T4 ligase (NEB) || 1.0 || 1.0 | ||
|- | |- | ||
| dH<sub>2</sub>O || | | dH<sub>2</sub>O || --- || 0.5 | ||
|- | |- | ||
| || | | || 10.0 μL || 10.0 μL | ||
|} | |} | ||
RESULTS (5/22/15) | |||
* Success! Ligation plates have 15-20 colonies, neg. ctrl has 1 colony | |||
* Pick 2 colonies from each plate for 5 mL cultures and streaks | |||
---- | |||
'''Ryan - pick pBra-BraR-MRV colonies" | |||
* pBra-BraR-MRV is the only receiver with no potential successes so far | |||
* Pick 8 colonies form the traditional ligation plate for 5 mL cultures (o/n) and streaks | |||
Latest revision as of 00:58, 27 September 2017
Karmella's BioBrick Cloning | Main project page Previous entry Next entry | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
05/21/15
Cas-tone Project
Thermal cycler: LabNet OptiMax "AnOlig shrt"
RESULTS (5/22/15)
Ryan - pick pBra-BraR-MRV colonies"
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