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July 18
 Colonies 1-6 from Gibson plates; 7 and 8 are reswabs of two colonies. From left to right: DNA ladder,(PNT001; colony 1; vector), P2v, P3v, P4v, P5v, P6v, P7v, P8v, (PNT001; colony 1; insert), P3i, P4i, P5i, P6i  from left to right: DNA ladder, (PNT001; reswab 7; insert), P8i, (16s; colony A; vector), 16Bv, 16Cv, (16s; A; insert), 16Bi, 16Ci
Colony PCR of 16s plasmids, for vector and insert
Gel of short PCR parts to confirm they are being amplified
Pulsed field gel with a larger amount of soil sample DNA
Retransform pSB3K5 into Xl-10s
Results
Gels show mostly expected single bands
July 19
PCR parts for Gibson assembly of pNT001, pNT003 and Joe's positive GFP control
Results
pSB3K5 plate had 0 colonies.
PFGE of soil-extracted DNA shows a smear; will do gel extraction of ~40kbp sequences for fosmid kit.
 lane 1 lambda mono-cut ladder, 2 blank, 3 40 kb control DNA, 4 LA River sample 9, 5 LA-River sample 10 Gibson PCR concentrations
| Part |
Concentration (ng/ul) |
| R0010 for pNT001 |
86.4 |
| B0014 for pNT001 |
128.3 |
| B0014 for pNT003 |
89.1 |
| promoter for GFP Gibson positive control |
51.9 |
| GFP for positive control |
130.4 |
| terminator for positive control |
82.5 |
July 20
Pulse field gel of soil-extracted DNA, in preparation for fosmid insertion
Redo Gibson assembly of pNT001 using a smaller total mass of DNA. Gibson assemble GFP from Joe that is known to have worked before as a positive control
July 21
Results
| Plate |
Number of Colonies |
| GFP + |
10 |
| GFP - |
3 |
| pNT001 + |
29 |
| pNT001 - |
2 |
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