"

From 2010.igem.org

Contents 1 Andreas 1.1 Removal of insertion in BioBrick suffixes 1.1.1 Transformation results 1.1.2 Colony PCR 1.1.3 Gel verification

Andreas

Removal of insertion in BioBrick suffixes

Transformation results

Good colony yield on all plates. A few red colonies discovered on all plates, indicating that some partly digested pSB1C3 vector, still containing the RFP insert, had been excised during gel extraction.

Colony PCR

From 11/10 transformations'

1. pSB1C3.IgGp A-C
2. pSB1C3.bFGF A-C
3. pSB1C3.ProtA A-C
4. pSB1C3.yCCS A-C
5. pSB1C3.SOD A-C

• Standard colony PCR settings
  • 1:15 elongation time

Gel verification

Gel 1: 1 % agarose, 140 V
Gel 2: 1 % agarose, 160 V

Expected bands

1. pSB1C3.IgGp: 1262 bp
2. pSB1C3.bFGF: 794 bp
3. pSB1C3.ProtA: 506 bp
4. pSB1C3.yCCS: 1076 bp
5. pSB1C3.SOD: 791 bp

Results

1. All clones correct.
2. All clones correct.
3. All clones correct.
4. Correct size for clone C, weak band for clone B. Clone A probably carrying an RFP insert.
5. All clones correct.