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From 2010.igem.org

Contents 1 Andreas 1.1 Assembly and cloning 1.1.1 Gel verification 1.1.2 ON cultures

Andreas

Assembly and cloning

Gel verification

Re-run of 27/9 colony PCRs.

Gel 1
1 % agarose, 110 V

Gel 2
0.8 % agarose, 90 V

Expected bands

1. 744 bp
2. 735 bp
3. 765 bp
4. 744 bp
5. 1645 bp
6. 1636 bp
7. 1666 bp
8. 1645 bp
9. 735 bp
10. 735 bp

Results
Listing clones of seemingly correct sizes, thereby potentially correct constructs:

1. 1
2. 1 & 2
3. 1 & 2
4. No correct clones
5. 2, 3 & 4
6. 1, 2 & 3
7. 1, 2, 3 & 4
8. No correct clones
9. 1 & 2
10. No correct clones

For further verification constructs should be sent for sequencing. However, we will await the sequencing results from samples sent 27/9, as they were also used for building these new assemblies.

ON cultures

Set ON cultures for plasmid prep (constructs 1-8) and glycerol stock (9)

• 5 ml LB + appr. antibiotic (Amp 100 or Km 50); 37 °C, 220 rpm
  1. 1
  2. 1
  3. 1
  4. -
  5. 2 & 3
  6. 2 & 3
  7. 1 & 2
  8. -
• 3 ml LB + Amp 100; 30 °C
  1. 1