User:Karmella Haynes/Notebook/BioBrick cloning/2013/04/01: Difference between revisions
From OpenWetWare
(Autocreate 2013/04/01 Entry for User:Karmella_Haynes/Notebook/BioBrick_cloning) |
(fix raw html notebook nav) |
||
(5 intermediate revisions by one other user not shown) | |||
Line 2: | Line 2: | ||
|- | |- | ||
|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}}}}]]}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
== | ==04/01/13== | ||
<!-- Precede finished items with a checkmark ✓ --> | <!-- Precede finished items with a checkmark ✓ --> | ||
* | * Check MV9 clones | ||
---- | ---- | ||
'''MV9 clones''' | |||
* Picked colonies from last cloning attempt. Streaked on LB amp agar. | |||
* Grew up 2 mL cultures overnight and did minipreps. | |||
'''Minipreps'''<br> | '''Minipreps'''<br> | ||
* Check with | * Check with S/X digests | ||
** S present in upstream cloning sites, X inside the new insert | |||
{| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | {| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | ||
Line 20: | Line 26: | ||
| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Volume | | bgcolor=#cfcfcf | Volume | ||
| rowspan="7" | <u>Expected:</u><br>1. | | rowspan="7" | <u>Expected:</u><br>1-4. MV9 = 4531, 80 | ||
| rowspan="7" | | | rowspan="7" | [[Image:KAH040113_gel1.jpg|200px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | ||
|- | |- | ||
| DNA(plasmid) || | | DNA(plasmid) || 3.0 μL | ||
|- | |- | ||
| 10X buffer || 1.5 | | 10X buffer || 1.5 | ||
Line 31: | Line 37: | ||
| PstI || 1.0 | | PstI || 1.0 | ||
|- | |- | ||
| dH<sub>2</sub>O || | | dH<sub>2</sub>O || 8.5 | ||
|- | |- | ||
| || 15 μL --> 37°C/ ~15 min. | | || 15 μL --> 37°C/ ~15 min. | ||
|} | |} | ||
* Result is unclear for #1. Submit for sequencing. | |||
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> |
Latest revision as of 22:35, 26 September 2017
Karmella's BioBrick Cloning | Main project page Previous entry Next entry | ||||||||||||||||
04/01/13
MV9 clones
|