Haynes Lab:Notebook/Engineering PC-TFs/2014/06/09: Difference between revisions
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==Summary== | ==Summary== | ||
'''Digest All the Things'''<br> | |||
---- | |||
'''Well One: Ladder'''<br> | |||
'''Well Two'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. Unknown <br> | |||
|- | |||
| DNA(CMV/MV9) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 0.0 μL | |||
|- | |||
| NruI || 0.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 8.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br> | |||
'''Well Three'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. 5200bp <br> | |||
|- | |||
| DNA(CMV/MV9) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| NruI || 0.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|}<br> | |||
'''Well Four'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. 5200bp <br> | |||
|- | |||
| DNA(CMV/MV9) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 0.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br> | |||
'''Well Five'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. 710bp, 4490bp <br> | |||
|- | |||
| DNA(CMV/MV9) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 6.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well Six'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. BL01 = 2520bp, V0120-2520bp <br> | |||
|- | |||
| DNA(BL01A) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| SpeI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well Seven'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. BL01 = 2520bp, V0120-2520bp <br> | |||
|- | |||
| DNA(BL01B) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| SpeI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well Eight'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. [[User:Cameron Lee Gardner|Cameron Lee Gardner]] 18:00, 10 June 2014 (EDT) <br> | |||
|- | |||
| DNA(BL05) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| SpeI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well Nine'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. [[User:Cameron Lee Gardner|Cameron Lee Gardner]] <br> | |||
|- | |||
| DNA(BL09) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| SpeI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well 10'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. BL01 = 7720bp <br> | |||
|- | |||
| DNA(CMV/MV9 & BL01) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 0.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 7.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br>'''Well 11'''<br> | |||
{| {{table}} border="1" cellspacing="3" | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. dependent on ligation direction <br> | |||
|- | |||
| DNA(CMV/MV9 & BL01) || 5.0 μL | |||
|- | |||
| 10X buffer || 1.5 μL | |||
|- | |||
| XbaI || 1.0 μL | |||
|- | |||
| NruI || 1.0 μL | |||
|- | |||
| dH<sub>2</sub>O || 6.5 μL | |||
|- | |||
| Total || 15 μL --> 37°C/ 30 min. | |||
|} | |||
<br> | |||
---- | |||
'''Gel Verification''' | |||
{| {{table}} border="1" cellspacing="3" <!-- Plates. table --> | |||
|- valign="top" | |||
| rowspan="7" | [[Image:20140609 172345.jpg|400px|Hover name]] <br> | |||
|} | |||
<!-- ##### 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. ##### --> |
Revision as of 15:00, 10 June 2014
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SummaryDigest All the Things Well One: Ladder
Well Four
Gel Verification
|