Ann's Notebook

From 2009.igem.org

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*Nanodrop of digest for Pu-RBS-GFP
*Nanodrop of digest for Pu-RBS-GFP
-
[[media:8_12_09_Digest_Pu_round_3_DNA_purification_and_nanodrop_of_Pu_GFP_digests.pdf| 8/12/09]]
+
[[media:8_13_09_Pu_GFP_leu_LP_preparations_for_ligation.pdf| 8/13/09]]
*Gel of purified DNA for digest round 1
*Gel of purified DNA for digest round 1
*Digest of DNA for Pu-RBS-GFP ligation round 2
*Digest of DNA for Pu-RBS-GFP ligation round 2
*Trouble shooting DNA purification process
*Trouble shooting DNA purification process
 +
*Gel of digested Pu, GFP, leu LP round 2
 +
*DNA purification of digested Pu, GFP, leu LP round 2
 +
*Nanodrop of digested Pu, GFP, leu LP round 2
 +
 +
GRE/VACATION
[[media:9_8_09_Pu_GFP_leu_landing_pad_ligation.pdf| 9/8/09]]
[[media:9_8_09_Pu_GFP_leu_landing_pad_ligation.pdf| 9/8/09]]
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*Start overnight cultures for Pu GFP leu landing pad miniprep
*Start overnight cultures for Pu GFP leu landing pad miniprep
-
[[media:9_12_09_Pu_GFP_leu_transformation_results.pdf| 9/13/09]]
+
[[media:9_22_09_Team_Meeting_Project_modifications.pdf| 9/13/09]]
*Pu GFP leu landing pad transformation results
*Pu GFP leu landing pad transformation results
*Start overnight cultures for Pu GFP leu landing pad miniprep
*Start overnight cultures for Pu GFP leu landing pad miniprep
Line 216: Line 221:
[[media:9_18_09_Primer_Design_for_Sequencing_of_Pu_GFP_leu.pdf| 9/18/09]]
[[media:9_18_09_Primer_Design_for_Sequencing_of_Pu_GFP_leu.pdf| 9/18/09]]
*Primer design for sequencing of Pu GFP leu landing pad
*Primer design for sequencing of Pu GFP leu landing pad
 +
 +
[[media:9_22_09_Team_Meeting_Project_modifications.pdf| 9/22/09]]
 +
*Weekly team meeting: project modifications
[[media:10_2_09_Pr_XylR_Primers.pdf| 10/2/09]]
[[media:10_2_09_Pr_XylR_Primers.pdf| 10/2/09]]
Line 249: Line 257:
*Gel of Digested Revised Pu, GFP generator, PrXylR, LeuLP and uncut Leu LP for ligation
*Gel of Digested Revised Pu, GFP generator, PrXylR, LeuLP and uncut Leu LP for ligation
*Ligation of Pu/GFP and PrXylR/LeuLP
*Ligation of Pu/GFP and PrXylR/LeuLP
 +
 +
[[media:10_15_09_10_16_09_Transformation_of_PrXylR_and_Pu_GFP_ligation.pdf| 10/15/09 abd 10/16/09]]
 +
*Comp. cell prep and transformation of PrXylR/LeuLP and Pu/GFP
 +
 +
[[media:10_18_09_Miniprep_of_Pu_GFP_ligation_round_2_and_PrXylR_ligation.pdf| 10/18/09]]
 +
*Miniprep of Pu/GFP and 1 PrXylR transformation colony
 +
 +
[[media:10_19_09_Digest_and_gel_of_miniprepped_PuGFP_PrLeuLP_and_prep_for_submission_into_biobricks2.pdf| 10/19/09]]
 +
*Nanodrop of miniprepped Pu/GFP and PrXylR/LeuLP ligation products
 +
*Digestion of miniprepped Pu/GFP and PrXylR/LeuLP ligation products
 +
*Gel of digested Pu/GFP and PrXylR/LeuLP
 +
*Submitting Parts to the Registry

Latest revision as of 03:08, 22 October 2009


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HOME THE TEAM THE PROJECT MODELING REGISTRY PARTS NOTEBOOK SAFETY



Ann's Notebook


6/19/09 and 6/25/09

  • M9 salts
  • Liquid LB
  • Antibiotic stock solutions
  • LB plates

7/22/09 and 7/23/09

  • Media preparation for transformation
  • Comp. cell protocol
  • Protocol for getting parts from the biobricks
  • Transformation protocol
  • Transformation of RBS for Pu promoter, Bacteriophage lysis cassette, lambda lysis device, T4 lysis device

7/24/09

  • Protocol for miniprep
  • Miniprep of RBS for Pu promoter, Bacteriophage lysis cassette, lambda lysis device, T4 lysis device

7/25/09

  • Protocol for using the nanodrop
  • Protocol for a digestion
  • Nanodrop concentrations of the RBS for Pu promoter, Bacteriophage lysis cassette, lambda lysis device, T4 lysis device
  • Digestion of the RBS for Pu promoter and the Bacteriophage lysis cassette

7/27/09

  • Plated overnight cultures for Pu colony PCR
  • Protocol for running a gel
  • Ran gel of the RBS for Pu promoter and Bacteriophage lysis cassette
  • Plated Pu-luciferase part sent from the registry

7/28/09

  • Overnight culture for frozen stock of Pu-luciferase biobrick
  • Digest of lambda and T4 killer genes to check part length with EcoRI, PstI, BglII and EcoRI/BglII

7/29/09

  • Frozen stock of Pu-luciferase biobrick
  • Gel of lambda and T4 killer genes digested with EcoRI, PstI, BglII and EcoRI/BglII
  • Digest of lambda and T4 killer genes with EcoRI/PstI, EcoRI/XhoI, XhoI

7/30/09

  • Gel of lambda and T4 killer genes digested with EcoRI/PstI, EcoRI/XhoI, XhoI
  • Gel of Pu promoter colony PCR

8/3/09

  • Analysis of Pu promoter colony PCR
    • Primer sequence
    • PCR conditions

8/4/09

  • Protocol for colony PCR
  • Colony PCR for Pu Promoter
  • Gel of Colony PCR for Pu promoter with samples at different annealing temperatures
  • Meeting with Pablo about HPLC

8/5/09

  • Protocol for DNA purification
  • DNA purification of Pu promoter colony PCR
  • Optimization of Pu promoter colony PCR
  • Nanodrop of Pu promoter

8/6/09

  • Protocol for DNA precipitation
  • DNA concentration of Pu promoter
  • Nanodrop results for concentrated Pu promoter

8/8/09

  • Comp. cell preparation and transformation of a GFP generator

8/9/09

  • Pu promoter colony PCR round 2
  • Gel of Pu promoter colony PCR round 2
  • DNA purification of colony PCR round 2
  • Nanodrop of colony PCR round 2 and leu landing pad
  • Miniprep of GFP generator
  • Nanodrop of GFP generator
  • Digest for Pu-RBS-GFP

8/10/09

  • Gel of digest for Pu-RBS-GFP

8/11/09

  • Digest of GFP generator
  • Gel of GFP generator
  • Pu promoter colony PCR round 3
  • Gel of Pu promoter colony PCR round 3
  • DNA purification of Pu promoter colony PCR round 3
  • Nanodrop of Pu promoter colony PCR round 3

8/12/09

  • Pu promoter round 3 digestion
  • DNA purification of digest for Pu-RBS-GFP
  • Nanodrop of digest for Pu-RBS-GFP

8/13/09

  • Gel of purified DNA for digest round 1
  • Digest of DNA for Pu-RBS-GFP ligation round 2
  • Trouble shooting DNA purification process
  • Gel of digested Pu, GFP, leu LP round 2
  • DNA purification of digested Pu, GFP, leu LP round 2
  • Nanodrop of digested Pu, GFP, leu LP round 2

GRE/VACATION

9/8/09

  • ligation protocol
  • Pu-GFP-leu landing pad ligation

9/9/09

  • ATCC sodium benzoate plate and liquid media recipes
  • Media prep for transformation of Pu GFP leu landing pad

9/10/09 and 9/11/09

  • Comp. cell preparation and transformation of Pu GFP leu landing pad ligation into E. coli and P. putida mt-2 (with TOL plasmid)

9/12/09

  • Pu GFP leu landing pad transformation results
  • Start overnight cultures for Pu GFP leu landing pad miniprep

9/13/09

  • Pu GFP leu landing pad transformation results
  • Start overnight cultures for Pu GFP leu landing pad miniprep

9/14/09

  • Nanodrop of Pu GFP leu landing pad miniprep
  • Digest of Pu GFP leu landing pad with EcoRI to determine the plasmids with the insert

9/15/09

  • Gel of Pu GFP leu landing pad ligation miniprepped products digested with EcoRI

9/16/09

  • Gel of Pu GFP leu landing pad ligation miniprepped products digested with EcoRI round 2

9/18/09

  • Primer design for sequencing of Pu GFP leu landing pad

9/22/09

  • Weekly team meeting: project modifications

10/2/09

  • Protocol for primer design
  • Primer design for PrXylR

10/5/09

  • Colony PCR PrXylR

10/6/09

  • Gel of Colony PCR PrXylR

10/9/09

  • Protocol for gel extraction
  • Gel extraction of PrXylR colony PCR
  • Nanodrop of PrXylR gel extraction
  • Gel of PrXylR gel extraction

10/10/09

  • PrXylR PCR amplification from gel extraction
  • Revised Pu promoter colony PCR
  • Gel of PrXylR and Revised Pu promoter PCR

10/11/09

  • Purification of revised Pu promoter
  • Nanodrop of revised Pu promoter
  • Digest of revised Pu promoter and GFP generator

10/13/09 abd 10/14/09

  • Miniprep of leu landing pad
  • Nanodrop of leu landing pad miniprep
  • Digest of leu landing pad and PrXylR
  • Gel of Digested Revised Pu, GFP generator, PrXylR, LeuLP and uncut Leu LP for ligation
  • Ligation of Pu/GFP and PrXylR/LeuLP

10/15/09 abd 10/16/09

  • Comp. cell prep and transformation of PrXylR/LeuLP and Pu/GFP

10/18/09

  • Miniprep of Pu/GFP and 1 PrXylR transformation colony

10/19/09

  • Nanodrop of miniprepped Pu/GFP and PrXylR/LeuLP ligation products
  • Digestion of miniprepped Pu/GFP and PrXylR/LeuLP ligation products
  • Gel of digested Pu/GFP and PrXylR/LeuLP
  • Submitting Parts to the Registry
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