User:Karmella Haynes/Notebook/BioBrick cloning/2012/12/11: Difference between revisions
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---- | ---- | ||
'''Assemblies''' | '''Assemblies''' | ||
# KAH111/C-His: 5' part/(E/P)/ | # KAH111/C-His: 5' part/(E/P)/1123 + C-His/(E/P)/3600 | ||
* Digests (Fermentas FD) | * Digests (Fermentas FD) | ||
{| {{table}} cellspacing="3" <!-- Digest rxn. table --> | {| {{table}} cellspacing="3" <!-- Digest rxn. table --> | ||
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| Sample || OD260 || 260/280 || ng/μL | | Sample || OD260 || 260/280 || ng/μL | ||
|- | |- | ||
| 1. KAH11 (E/P) || | | 1. KAH11 (E/P) || 0.019 || 1.96 || 18.7 | ||
|- | |- | ||
| 2. C-His (E/P) || | | 2. C-His (E/P) || 0.047 || 1.82 || 46.8 | ||
|} | |} | ||
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| Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> 12/12/12</font> | | Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> 12/12/12</font> | ||
|- | |- | ||
| 1. KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng || <font color="blue"> | | 1. KAH111(E/P)/size, 16 ng + C-His(E/P)/3600, 25ng || <font color="blue">KAH111/C-His 3:1 (Pick 4)</font> (very small colonies after ~18 hours) | ||
|- | |- | ||
| 2. C-His(E/P)/ 25 ng || | | 2. C-His(E/P)/ 25 ng || 3 colonies | ||
|} | |} | ||
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* pSB1A3-RFP (retransformation from 12/10/12) | * pSB1A3-RFP (retransformation from 12/10/12) | ||
* Note: pellets are red | * Note: pellets are red | ||
* Digest 10 μL of each with X/S for Gibson cloning (Brady and Vi) | * Digest 10 μL of each with X/S for Gibson cloning (Brady and Vi) | ||
* Note: combined mini preps afterwards; concentration = 60 ng/μL | |||
{| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | {| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | ||
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| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| rowspan="7" | <u>Expected:</u><br>3. pSB1A3 = | | rowspan="7" | <u>Expected:</u><br>3. pSB1A3 = 2155<br>4. pSB1A3 = 2155<br> | ||
| rowspan="7" | [[Image:KAH121112_gel1.png|200px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | | rowspan="7" | [[Image:KAH121112_gel1.png|200px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
|- | |- | ||
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* Gel purification: combined the two slices | * Gel purification: combined the two slices | ||
** Concentration: 260 = , 260/280 = , '''ng/μL = ''' | ** Concentration: 260 = 0.015, 260/280 = 2.2, '''ng/μL = 15.1''' | ||
Revision as of 11:27, 12 December 2012
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12/11/12
Assemblies
Minipreps
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