Team:UNIPV-Pavia/Calendar/July/settimana2

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Latest revision as of 16:53, 24 October 2010

JULY: WEEK 2

July, 5th

LB agar plates prepared:

LB+Amp 100 (500ml)
LB+Cm 34 (500ml)
LB (250ml)
LB+Cm 12.5 (250ml)

^top

July, 6th

We decided to perform a TECAN test to evaluate the strength of some promoters belonging to the Anderson Promoters Collection. Inoculum from glycerol stocks (8ul in 5ml LB) for:

<partinfo>BBa_J23101</partinfo>
<partinfo>BBa_J23110</partinfo>
<partinfo>BBa_J23116</partinfo>
<partinfo>BBa_J23118</partinfo>
<partinfo>BBa_J23114</partinfo>
<partinfo>BBa_B0033</partinfo>

TECAN experiment started at 10:20am.

In the morning one single colony was picked for BW53474 E. coli strain and glycerol stock was prepared after about 7 hours (37°C 220rpm)

Inoculum of:

BW53474 (falcon containing culture for glycerol stock was re-filled with 5ml LB)
MG1655 (from glycerol stock)
BW25141 (from glycerol stock)
BW25142 (from glycerol stock)

^top

July, 7th

Competent cells preparation for:

MG1655
BW53474
BW25141
BW25142

PCR was performed to amplify Phasins:

All parts were correct!! :)

^top

July, 8th

Transformation of 1ul (~4ng) of miniprepped <partinfo>BBa_J23118</partinfo> into 100ul of home-made competent cells

MG1655
BW25141
BW25142
BW53474
DH5-alpha (as control)

to verify the transformation efficiency.

Transformed bacteria were plated on LB+Amp and left overnight in oven, 37°C.

^top

July, 9th

We checked the presence of colonies in plates. All plates showed red colonies!

MG1655 plate (with satellite colonies)	BW25141 plate	BW25142 plate
BW53474 plate	DH5-alpha plate

We calculated (thanks Nicolò) efficiency as #colonies/ug DNA

Strain	#colonies	Efficiency
MG1655	379	~10^5
BW25141	1431	~10^5
BW25142	2374	~10^5
BW53474	9984	~10^6 (very difficult to count correctly)
DH5alpha	7475 (only 100ul of cells plated)	~ 10^6 (very difficult to count correctly, previous tests showed an efficiency of ~10^8)

^top

@@ Line 11: / Line 11: @@
 <td valign="top">
 <table border="0" align="center" width="100%"><tr><td align="justify" valign="top" style="padding:20px">
+<table class="menu" border="0" width="100%">
+<tr>
+<td align="center">
+[[Team:UNIPV-Pavia/Calendar/July/settimana1|Week 1]]
+</td>
+<td align="center" style="padding:0; height:20px">
+[[Team:UNIPV-Pavia/Calendar/July/settimana2|Week 2]]
+</td>
+<td align="center">
+[[Team:UNIPV-Pavia/Calendar/July/settimana3|Week 3]]
+</td>
+<td align="center">
+[[Team:UNIPV-Pavia/Calendar/July/settimana4|Week 4]]
+</td>
+<td align="center">
+[[Team:UNIPV-Pavia/Calendar/July/settimana5|Week 5]]
+</td>
+</tr>
+</table>
+<br>
 <html><p align="center"><font size="4"><b>JULY: WEEK 2</b></font></p></html><hr><br>
+<html><a name="indice"/></html>
 ==July, 5th==
@@ Line 19: / Line 42: @@
 *LB (250ml)
 *LB+Cm 12.5 (250ml)
+<div align="right"><small>[[#indice|^top]]</small></div>
 ==July, 6th==
-We decided to perform a <font size=4>[[Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test3|TECAN test]]</font> to evaluate the strength of some promoters belonging to the Anderson Promoters Collection. Inoculum from glycerol stpcks (8ul in 5ml LB) for:
+We decided to perform a TECAN test to evaluate the strength of some promoters belonging to the Anderson Promoters Collection. Inoculum from glycerol stocks (8ul in 5ml LB) for:
 *<partinfo>BBa_J23101</partinfo>
 *<partinfo>BBa_J23110</partinfo>
@@ Line 32: / Line 57: @@
 TECAN experiment started at 10:20am.
-In the morning one single colony was peaked for BW53474 E. coli strain and glycerol stock was prepared after about 7 hours (37°C 220rpm)
+In the morning one single colony was picked for BW53474 E. coli strain and glycerol stock was prepared after about 7 hours (37°C 220rpm)
 Inoculum of:
@@ Line 39: / Line 64: @@
 *BW25141 (from glycerol stock)
 *BW25142 (from glycerol stock)
+<div align="right"><small>[[#indice|^top]]</small></div>
 ==July, 7th==
-Competent cells preparationf for:
+Competent cells preparation for:
 *MG1655
 *BW53474
@@ Line 49: / Line 76: @@
 PCR was performed to amplify Phasins:
-[[Image:Phasins PCR.jpg|300px|thumb|center|Phasins amplified by PCR (Marker-Blank-PhaP1-PhaP1-PhaP2-PhaP2)]]
 All parts were correct!! :)
+<div align="right"><small>[[#indice|^top]]</small></div>
 ==July, 8th==
@@ Line 65: / Line 92: @@
 Transformed bacteria were plated on LB+Amp and left overnight in oven, 37°C.
+<div align="right"><small>[[#indice|^top]]</small></div>
 ==July, 9th==
@@ Line 93: / Line 122: @@
 |}
-<td width="20%" valign="top">
+<div align="right"><small>[[#indice|^top]]</small></div>
-<table border="0" width="100%" height="100%">
+<!-- table previous next week -->
-<tr align="right"><td height="15">
+<br><br>
-[[Team:UNIPV-Pavia/Calendar|Calendar]]
+<table border="0" width="100%" class="menu">
-</td></tr>
+<tr>
-<tr align="right"><td height="15">
+<td align="left">[[Team:UNIPV-Pavia/Calendar/July/settimana1| Previous week]]</td>
-[[Team:UNIPV-Pavia/Calendar/July|July]]
+<td align="right">[[Team:UNIPV-Pavia/Calendar/July/settimana3| Next week]]</td>
-</td></tr>
+</tr>
-<tr align="right"><td height="15">
-[[Team:UNIPV-Pavia/Calendar/July/settimana1|week 1]]
-</td></tr>
-<tr align="right"><td height="15">
-[[Team:UNIPV-Pavia/Calendar/July/settimana2|week 2]]
-</td></tr>
-<tr align="right"><td height="15">
-[[Team:UNIPV-Pavia/Calendar/July/settimana3|week 3]]
-</td></tr>
-<tr align="right"><td height="15">
-[[Team:UNIPV-Pavia/Calendar/July/settimana4|week 4]]
-</td></tr>
-<tr align="right"><td height="15">
-[[Team:UNIPV-Pavia/Calendar/July/settimana5|week 5]]
-</td></tr>
 </table>
+<!-- fine table previous next week -->
+</td>
+<td width="15%" align="right" valign="top">
+{{UNIPV-Pavia/menu_mesi}}
 </td>
 </tr>
 </table>