Team:Toronto/Notebook-w09-wed
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Wednesday July 13th
Wednesday, 7/13
Members Present: Alex, Cathy, Tam, Dk, Hamed
LAB:
Morning:
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A gel was ran on the previous day's miniprep results:
Lane 1: NEB 2 Log DNA Ladder (10kb)
Lane 2: GolS(W) - 1 band in between 1.5kb and 2kb
Lane 3: GolS(R) - 2 bands in between 2kb and 3kb
Lane 4: mCherry - 1 band in between 1.5kb and 2kb
Lane 5: P118A - 2 bands in between 2kb and 3kb
NOTE: There appears to be faded bands in lane 2 above the 10kb, lane 4 above 10kb and around 4kb and lane 5 around 6kb.
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Wells contained 8ul of PCR sample with 1.6ul of DNA Loading Dye
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Well 4 was the DNA Ladder: 2-log DNA Ladder (0.1 - 10.0 kb)
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Gel ran at 100V for 60 min
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Visualize using GeneSnap with Transilliminator and Et/UV
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600ml of 1x TAE was made and 600ml of autoclaved MilliQ water was prepared
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Disposed of:
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2 old LB media (both appeared to have less than 100ml in it
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Old bottle of SOC
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Dirty bottle of autoclaved MilliQ water
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Old bottle of autoclaved dH2O
Afternoon:
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Counted the colonies on each plate (from transformations done yesterday)
A | B | C | D | |
1 | Plate | Caption | A | B |
2 | 1 | TetO_LacZ | 6 | 9 |
3 | 2 | TetO_mCherry | 17 | 14 |
4 | 3 | pgoB_LacZ | 8 | 11 |
5 | 4 | Long_GolSP11A | 28 | 13 |
6 | 5 | Short_TetO_GolS (P118A) | 50+ | 70+ |
7 | 6 | Long_GolS | 6 | 10 |
8 | 7 | Short_TetO_GolS | 50+ | 100 (approx.) |
9 | 8 | Short_PrcnA_LacZ | 11 | 7 |
10 | 9 | Short_TetO_RcnR | 16 | 12 |
Table1
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Measured OD600 of the BL21 culture (it was 1.566 at 5 P.M.). We decided to grow them until tomorrow.
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Made LB+Agar+CAM and LB+Agar plates
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Did 18x5mL cultures for miniprep tomorrow (L1A, L1B, L2A, ..., L9B). They are in rm 315 shaker along with the BL21 cultures.
Administrative:
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Alex had meeting with Dennis from OG:
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No updates from Nathan Stubina from McEwen Mining Industries
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Marc Fume continues interest and has informed his gold mining contacts about us
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Alex will send follow-up soon
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Server for the P&P synthetic biology network was denied but they may be willing to give it to us if we provide them a letter that it is an oGEM initiative instead of iGEM Toronto initiative
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OG is still awaiting invoices from CSM for reimbursements
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University College has given us $500
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Purchases made for the day
A | B | C | D | E | F | |
1 | Name | SKU | Unit Price | Quantity | Total (incl. tax) | Status |
2 | VWR Petri dishes (100x15mm, 25pcs/case) | 25384-342 | 3.6 | 5 | 20.34 | Available |
3 | Falcon tubes (50mL, 15pcs/rack) | 352098 | 6.56 | 4 | 29.65 | Available |
4 | Liquid nitrogen | 2384 | 1.71 | 3 | 5.80 | Available |
5 | Grand Total | 55.79 | ||||
Table2
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PEP status update: shipping from Sigma USA on July 27th
TO DO:
For the next day:
LAB TEAM:
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Miniprep cultures that were grown on Wednesday, July 13th and run a gel on them
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Check OD of BL21 cultures and do the 3xwash and flash freezing steps
LAB MANAGERS:
