Haynes Lab:Notebook/Engineering PC-TFs/2014/06/16: Difference between revisions

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(Autocreate 2014/06/16 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs)
 
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==Summary==
==Summary==
*
'''Digest All the Things Addendum'''<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> 2520(insert)+4968(V0120 Vector) = 7488bp <br><u>Observed:</u><br> <br>
|-
| DNA(BL05 M.P) || 8.0 μL
|-
| 10X buffer || 1.5 μL
|-
| XbaI || 1.0 μL
|-
| dH<sub>2</sub>O || 4.5 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>2520(insert)+4968(V0120 Vector) = 7488bp <br><u>Observed:</u><br> <br>
|-
| DNA(BL05A) || 4.0 μL
|-
| 10X buffer || 1.5 μL
|-
| XbaI || 1.0 μL
|-
| dH<sub>2</sub>O || 8.5 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>1431 (insert) + 4968 (V0120 Vector) = 6399bp <br><u>Observed:</u><br> <br>
|-
| DNA(BL09 M.P.) || 8.0 μL
|-
| 10X buffer || 1.5 μL
|-
| XbaI || 1.0 μL
|-
| dH<sub>2</sub>O || 4.5 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> 1431 (insert) + 4968 (V0120 Vector) = 6399bp <br><u>Observed:</u><br> <br>
|-
| DNA(BL09B) || 4.0 μL
|-
| 10X buffer || 1.5 μL
|-
| XbaI || 1.0 μL
|-
| dH<sub>2</sub>O || 8.5 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> 5197bp <br><u>Observed:</u><br> <br>
|-
| DNA(Phosphorylated CMV/MV9 ) || 2.0 μL
|-
| 10X buffer || 1.5 μL
|-
| dH<sub>2</sub>O || 11.5 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> [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
|}
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Revision as of 14:54, 16 June 2014



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Summary

Digest All the Things Addendum

Well One: Ladder
Well Two

Reagent Volume Expected:
2520(insert)+4968(V0120 Vector) = 7488bp
Observed:

DNA(BL05 M.P) 8.0 μL
10X buffer 1.5 μL
XbaI 1.0 μL
dH2O 4.5 μL
Total 15 μL --> 37°C/ 30 min.


Well Three

Reagent Volume Expected:
2520(insert)+4968(V0120 Vector) = 7488bp
Observed:

DNA(BL05A) 4.0 μL
10X buffer 1.5 μL
XbaI 1.0 μL
dH2O 8.5 μL
Total 15 μL --> 37°C/ 30 min.

Well Four

Reagent Volume Expected:
1431 (insert) + 4968 (V0120 Vector) = 6399bp
Observed:

DNA(BL09 M.P.) 8.0 μL
10X buffer 1.5 μL
XbaI 1.0 μL
dH2O 4.5 μL
Total 15 μL --> 37°C/ 30 min.


Well Five

Reagent Volume Expected:
1431 (insert) + 4968 (V0120 Vector) = 6399bp
Observed:

DNA(BL09B) 4.0 μL
10X buffer 1.5 μL
XbaI 1.0 μL
dH2O 8.5 μL
Total 15 μL --> 37°C/ 30 min.


Well Six

Reagent Volume Expected:
5197bp
Observed:

DNA(Phosphorylated CMV/MV9 ) 2.0 μL
10X buffer 1.5 μL
dH2O 11.5 μL
Total 15 μL --> 37°C/ 30 min.


Gel Verification

Hover name
Ladder

|}


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