Team:Columbia-Cooper-NYC/Columbia notebook 2

From 2012.igem.org

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=== Week 4 ===
=== Week 4 ===
==== Tuesday, 24th ====
==== Tuesday, 24th ====
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*
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* Re-hydrated four biobrick parts according to standard iGEM re-hydration protocol
 +
*# Inducible plasmid (Bba J04500) in plasmid pSB1AK3 from plate 4, 12A
 +
*# GFP (Bba E0040) in plasmid pSB1A2 from plate 1, 14K
 +
*# High copy plasmid (Bba J044500) in plasmid pSB1T3 from plate 1, 7A
 +
*# Low copy plasmid (Bba J044500) in plasmid pSB3C5
==== Wednesday, 25th ====
==== Wednesday, 25th ====
==== Thursday, 26th ====
==== Thursday, 26th ====

Revision as of 01:53, 2 August 2012

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Columbia Genetics Lab Notebook

July, 2012

Week 1

Thursday, 5th

  • Re-hydrated plasmids with 50µl of LB and Kanamycin solution
  • Stored solution at 37°C incubator overnight

Friday, 6th

  • Purified pET26b vector using standard DNA purification protocol

Week 2

Monday, 9th

  • Received kill gene Bba-K124017 from plate 3, 20M
  • Re-hydrated DNA according to standard iGEM re-hydration protocol

Tuesday, 10th

  • Contacted professors at Germany in hopes to receive copies of fungal phytochrome FphA

Wednesday, 11th

  • Received confirmation by professors at Germany for FphA to be sent to Columbia University
  • Conducted transformation using electroporation with competent bacteria (marked by resistance to Kanamycin)
    1. Control: 1µl of deionized water with abt. and 60µl of bacteria cells
    2. Variable: 1µl of re-hydrated kill gene and 60µl of bacteria cells
  • Placed both samples after electroporation into 200µl of preprepared LB
  • Placed samples in shaker 37°C for 30 minutes

Thursday, 12th

  • Grew 1 colony of transformed bacteria in 5mL of Kanamycin and LB solution
    • Note: Using pET20b vector over pET26b vector from glycerol stock solution

Friday, 13th

  • Isolated 4 samples of plasmid using standard plasmid isolation protocol
    1. 2 samples: kill gene
    2. 2 samples: pET20b vector

Week 3

Monday, 16th

  • Re-hydrated two biobrick parts in plasmid pSB2K3 according to standard iGEM re-hydration protocol
    1. BBa-I16009 (PcyA) from plate 1, 20F
    2. BBa-I16008 (ho1) from plate 2, 13J
  • Electroporated 1µl of each biobrick into separate E. coli at 1800V
  • Added 100µl LB broth into each sample
  • Placed samples at 33.4°C for 20 minutes
  • Samples were plated to be grown overnight

Tuesday, 17th

  • Placed 5ml each of LB/Kan into two centrifuge tube for PCB creation
    1. Label P: PcyA
    2. Label h: ho1
  • Placed samples in 37°C incubator

Wednesday, 18th

  • Purified ho1 and PcyA plasmids using standard DNA purification protocol
  • Placed purified DNA into glycerol stock (LB/Kan) and stored at -80°C

Thursday, 19th

  • Purified GFP using standard DNA purification protocol
  • Prepared glycerol stock solution (500µl GFP/500µl 90% glycerol) and stored at -80°C

Week 4

Tuesday, 24th

  • Re-hydrated four biobrick parts according to standard iGEM re-hydration protocol
    1. Inducible plasmid (Bba J04500) in plasmid pSB1AK3 from plate 4, 12A
    2. GFP (Bba E0040) in plasmid pSB1A2 from plate 1, 14K
    3. High copy plasmid (Bba J044500) in plasmid pSB1T3 from plate 1, 7A
    4. Low copy plasmid (Bba J044500) in plasmid pSB3C5

Wednesday, 25th

Thursday, 26th

Friday, 27th

Saturday, 28th

Week 5

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