Glasgow/Wetlab/Week3

From 2007.igem.org

< Glasgow | Wetlab(Difference between revisions)
 
(13 intermediate revisions not shown)
Line 1: Line 1:
-
[[Glasgow|Glasgow Main Page]] | [[Glasgow/Wetlab|Back To Wetlab Log]]
+
{| valign=top cellpadding=3
 +
|-
 +
!align=center|[[Image:Uog.jpg]] ||  [[Glasgow|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Main Page</font>]] || [[Glasgow/Wetlab|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Wetlab Log</font>]] ||  [[Glasgow/Modeling|<font face=georgia color=#3366CC size=4>Go To <br> Glasgow's <br>Drylab Log</font>]]
 +
|}
----
----
{|cellspacing="6px" cellpadding="16" border="0" width="100%"
{|cellspacing="6px" cellpadding="16" border="0" width="100%"
-
|-
+
|- align=center
-
|[[Glasgow/Wetlab/Protocols|PROTOCOLS]]
+
|[https://2007.igem.org/Glasgow/Wetlab/Protocols <font face=georgia color=#3366CC size=5><b>Protocols</b></font>]
-
|[[Glasgow/Wetlab/References|REFERENCES]]
+
|[https://2007.igem.org/Glasgow/Wetlab/References <font face=georgia color=#3366CC size=5><b>References</b></font>]
-
|[[Glasgow/Wetlab/Resources|RESOURCES]]
+
|[https://2007.igem.org/Glasgow/Wetlab/Resources <font face=georgia color=#3366CC size=5><b>Resources</b></font>]
-
|[[Glasgow/Wetlab/Orders|ORDERS]]
+
|[https://2007.igem.org/Glasgow/Wetlab/Orders <font face=georgia color=#3366CC size=5><b>Orders</b></font>]
-
|[[Glasgow/Wetlab/Biobricks|BIOBRICKS UTILISED]]
+
|[https://2007.igem.org/Glasgow/Wetlab/Biobricks <font face=georgia color=#3366CC size=5><b>Biobricks<br>Used</b></font>]
-
|[[Glasgow/Wetlab/Gels|GELS]]
+
|[https://2007.igem.org/Glasgow/Wetlab/Gels <font face=georgia color=#3366CC size=5><b>Gels</b></font>]
-
 
+
|}
|}
----
----
Line 70: Line 72:
#Last night we diluted primers to 100 pmol/ul by adding the amount of dH2O given on the order sheet, and made working solutions to 10 pmol/ul (2 x 200 ul each - 20 ul stock and 180 ul dH2O).
#Last night we diluted primers to 100 pmol/ul by adding the amount of dH2O given on the order sheet, and made working solutions to 10 pmol/ul (2 x 200 ul each - 20 ul stock and 180 ul dH2O).
#[[User:MaijaP|Maija]] ran a gel of the restriction digests to check the sizes of the biobricks.  These were the maps we made yesterday.
#[[User:MaijaP|Maija]] ran a gel of the restriction digests to check the sizes of the biobricks.  These were the maps we made yesterday.
-
#[[User:Mojs|Maia]] is extracting DNA from 3 samples of ****, which were plated yesterday and grown in liquid media, using MoBio PowersoilTM DNA purification kit (see [[Glasgow/Wetlab/Protocols#Protocol 8: MoBio PowersoilTM DNA Purification Kit|Protocol 8]]); only change was to shorten "shaking" time from 10 to 2 minutes.
+
#[[User:Mojs|Maia]] is extracting DNA from 3 samples of Pseudomonas Aeruginosa, which were plated yesterday and grown in liquid media, using MoBio PowersoilTM DNA purification kit (see [[Glasgow/Wetlab/Protocols#Protocol 8: MoBio PowersoilTM DNA Purification Kit|Protocol 8]]); only change was to shorten "shaking" time from 10 to 2 minutes.
-
#PCR trial run using Reddymix and Touch 2 done (See [[Glasgow/Wetlab/Protocols#Protocol 9: PCR|Protocol 9]] for PCR).
+
#PCR trial run using Reddymix and Touch 2 done (see [[Glasgow/Wetlab/Protocols#Protocol 9: PCR|Protocol 9]] for PCR).
{| border="1" cellspacing="0" cellpadding="5" align="center"
{| border="1" cellspacing="0" cellpadding="5" align="center"
!Template
!Template
Line 92: Line 94:
=== Thursday 19th July 2007 ===
=== Thursday 19th July 2007 ===
-
#[[User:Mojs|Maia]] redid PCR on pGLTUR and pQF52 using Reddymix and a gradient program (See [[Glasgow/Wetlab/Protocols#Protocol 9: PCR|Protocol 9]]) to minimise the non-specific amplification evident in previous gels. New DntR suffix primer has arrived. Used the following primer sets.
+
#[[User:Mojs|Maia]] redid PCR on pGLTUR and pQF52 using Reddymix and a gradient program (see [[Glasgow/Wetlab/Protocols#Protocol 9: PCR|Protocol 9]]) to minimise the non-specific amplification evident in previous gels. New DntR suffix primer has arrived. Used the following primer sets.
#*XylR prefix and XylR suffix
#*XylR prefix and XylR suffix
#*Pr prefix and Pr suffix
#*Pr prefix and Pr suffix
Line 100: Line 102:
=== Friday 20th July 2007 ===
=== Friday 20th July 2007 ===
 +
 +
<br>
 +
{| valign=top cellpadding=3
 +
|-
 +
![[Glasgow/Wetlab/Week2|<font face=georgia color=#3366CC size=4>Previous  <br>  Week</font>]] || [[Glasgow/Wetlab/Week4|<font face=georgia color=#3366CC size=4>Next <br>  Week</font>]]
 +
|-
 +
|}
 +
----
 +
{| valign=top cellpadding=3
 +
|-
 +
!align=center|[[Image:Uog.jpg]] ||  [[Glasgow|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Main Page</font>]] || [[Glasgow/Wetlab|<font face=georgia color=#3366CC size=4>Back To <br> Glasgow's <br> Wetlab Log</font>]]
 +
|}

Latest revision as of 17:08, 22 October 2007

Uog.jpg Back To
Glasgow's
Main Page 		Back To
Glasgow's
Wetlab Log 		Go To
Glasgow's
Drylab Log
Protocols References Resources Orders Biobricks
Used 		Gels

Contents

Week 3

Tuesday 17th July 2007

  1. Restriction Digests of transformed BioBricks (see Protocol 7):
Label BioBrick Plasmid Description Enzymes Expected Sizes
3/19A BBa_J23119 pSB1A2 strong constitutive promoter NheI, PvuI 680bp, 1430bp
1/9G BBa_R0062 pBB1A2 HSL and luxR EcoRI, PvuI 1460bp, 660bp
1/16P BBa_J04500 pSB1AK3 IPTG inducer and RBS PvuII, PvuI 2250bp, 1030bp, 730bp
4/11C BBa_p1010 pSB3K3 death gene BamHI, XhoI 190bp, 2390bp,840bp
1/5H BBa_E0040 pSB1A2 GFP no promoter Hime II, PvuI 1630bp, 1170bp
4/6D BBa_I52001 p5B4K5 High copy death gene AvaI, PvuI 920bp, 1470bp, 2120bp

Wednesday 18th July 2007

  1. Last night we diluted primers to 100 pmol/ul by adding the amount of dH2O given on the order sheet, and made working solutions to 10 pmol/ul (2 x 200 ul each - 20 ul stock and 180 ul dH2O).
  2. Maija ran a gel of the restriction digests to check the sizes of the biobricks. These were the maps we made yesterday.
  3. Maia is extracting DNA from 3 samples of Pseudomonas Aeruginosa, which were plated yesterday and grown in liquid media, using MoBio PowersoilTM DNA purification kit (see Protocol 8); only change was to shorten "shaking" time from 10 to 2 minutes.
  4. PCR trial run using Reddymix and Touch 2 done (see Protocol 9 for PCR).
Template Primers
pGLTUR


XylR_prefix + XylR_suffix
Pr_prefix + Pr_suffix
Pr_prefix + XylR_suffix
Pu_prefix_EM + Pu_suffix_EM

pQF52 DntR_prefix + DntR_suffix
DmpR WT/24 DntR_prefix + DntR_suffix

Thursday 19th July 2007

  1. Maia redid PCR on pGLTUR and pQF52 using Reddymix and a gradient program (see Protocol 9) to minimise the non-specific amplification evident in previous gels. New DntR suffix primer has arrived. Used the following primer sets.
    • XylR prefix and XylR suffix
    • Pr prefix and Pr suffix
    • Pr prefix and XylR suffix
    • Pu prefix and Pu suffix
    • DntR prefix and DntR suffix 2

Friday 20th July 2007


Previous
Week 		Next
Week
Uog.jpg Back To
Glasgow's
Main Page 		Back To
Glasgow's
Wetlab Log
Retrieved from "http://2007.igem.org/wiki/index.php/Glasgow/Wetlab/Week3"