Glasgow/Wetlab/Week3

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[[Glasgow|Glasgow Main Page]]  |  [[Glasgow/Wetlab|Back To Wetlab Log]]
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== Week 3 ==
== Week 3 ==
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=== Tuesday 17/07 ===
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=== Tuesday 17th July 2007 ===
#Restriction Digests (see [[Glasgow/Wetlab/Protocols#Protocol 7: Restriction Digests|Protocol 7]]):
#Restriction Digests (see [[Glasgow/Wetlab/Protocols#Protocol 7: Restriction Digests|Protocol 7]]):
#*BBa_J23119 (strong constitutive promoter) pSB1A2.  1 x NheI, 1 x PvuI.  680bp, 1430bp
#*BBa_J23119 (strong constitutive promoter) pSB1A2.  1 x NheI, 1 x PvuI.  680bp, 1430bp
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#*BBa_I52001 (6D) p5B4K5. 2 x AvaI, 1 x PvuI. 920bp, 1470bp, 2120bp.
#*BBa_I52001 (6D) p5B4K5. 2 x AvaI, 1 x PvuI. 920bp, 1470bp, 2120bp.
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=== Wednesday 18/07 ===
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=== Wednesday 18th July 2007 ===
#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).
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#Maija ran a gel of the restriction digests to check the sizes of the biobricks.  These were the maps we made yesterday.
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#[[User:MaijaP|Maija]] ran a gel of the restriction digests to check the sizes of the biobricks.  These were the maps we made yesterday.
-
#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.
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#[[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.
#PCR trial run using Reddymix and Touch 2 done on:
#PCR trial run using Reddymix and Touch 2 done on:
#*pGLTUR: XylR_prefix / XylR_suffix, Pr_prefix / Pr_suffix, Pr_prefix / XylR_suffix, Pu_prefix_EM / Pu_suffix_EM.
#*pGLTUR: XylR_prefix / XylR_suffix, Pr_prefix / Pr_suffix, Pr_prefix / XylR_suffix, Pu_prefix_EM / Pu_suffix_EM.
#*pQF52 plasmid: DntR_prefix / DntR_suffix
#*pQF52 plasmid: DntR_prefix / DntR_suffix
#*DmpR WT/24: DntR_prefix / DntR_suffix
#*DmpR WT/24: DntR_prefix / DntR_suffix
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#*Biobricks: R0062, E0040, J04500, J23119, p1010, I52001.
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#*Biobricks: [http://partsregistry.org/Part:BBa_R0062 R0062], [http://partsregistry.org/Part:BBa_E0040 E0040], [http://partsregistry.org/Part:BBa_J04500 J04500], [http://partsregistry.org/Part:BBa_J23119 J23119], http://partsregistry.org/Part:BBa_p1010 p1010], http://partsregistry.org/Part:BBa_I52001 I52001].
#*See [[Glasgow/Wetlab/Protocols#Protocol 9: PCR|Protocol 9]] for PCR.
#*See [[Glasgow/Wetlab/Protocols#Protocol 9: PCR|Protocol 9]] for PCR.
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=== Thursday 19/07 ===
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=== Thursday 19th July 2007 ===
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#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.
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#[[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
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#*DntR prefix and DntR suffix 2
#*DntR prefix and DntR suffix 2
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=== Friday 20/07 ===
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=== Friday 20th July 2007 ===

Revision as of 18:33, 15 August 2007

Glasgow Main Page | Back To Wetlab Log

Contents

Week 3

Tuesday 17th July 2007

  1. Restriction Digests (see Protocol 7):
    • BBa_J23119 (strong constitutive promoter) pSB1A2. 1 x NheI, 1 x PvuI. 680bp, 1430bp
    • BBa_R0062 (HSL and luxR) pBB1A2. 1 x EcoRI, 1 x PvuI. 1460bp, 660bp
    • BBa_J04500 (IPTG inducer and RBS) pSB1AK3. 1 x PvuII, 2 x PvuI. 2250bp, 1030bp, 730bp
    • BBa_p1010 (death gene) pSB3K3. 1 x BamHI, 2 x XhoI. 190bp, 2390bp,840bp.
    • BBa_E0040 (GFP no promoter) pSB1A2. 1 x Hime II, 1 x PvuI. 1630bp, 1170bp.
    • BBa_I52001 (6D) p5B4K5. 2 x AvaI, 1 x 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 ****, 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 on:

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

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