Team:Stanford/NotebookPage/8 July 2010
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Christopher's Notebook Section
- Running a Gel of F2620, pBAD, and psb4A5:
- Make a 0.8% agarose gel (0.4 g of agarose in 50 ml of TAE)
- Heat in microwave until agarose is completely dissolved
- Add 10.0 ul of EtBr to mix (5000x for a 50 ml solution)
- let run for approximately 1 hr
- refer below for bands
4A5 E/P 3395 bp I0500 E/S (on psb2K3) 1210 bp F2620 E/S (on pSB1A2) 1061 bp
Gel Extract the Fragments Running a Gel of PCR Product: -first need to PCR cleanup -then run on 1.4% agarose gel (0.7 g of agarose in 50 ml of TAE) -this is necessary because of the 800 bp size of the fragment
Miniprep Ligations -F2620+E0240+psb1A2 -pBAD+E0240+psb1A2
Restriction Digests PCR product F2620
Laura's Lab notebook
miniprepped Alex's/Chris' liquid cultures, using Promega kit
- 2 each of:
- psb1K3
- J23107
- B0032
- eluted in 100 uL H2O
Nanodrop Data | |||
---|---|---|---|
tube ID | 260/280 | 260/230 | ng/uL |
psB1K3-1 | 1.77 | 1.08 | 71.3 |
psB1K3-2 | 1.73 | 1.01 | 77.8 |
J23107-1 | 1.87 | 1.60 | 116.3 |
J23107-2 | 1.90 | 1.74 | 142.0 |
B0032-1 | 1.81 | 1.20 | 68.1 |
B0032-2 | 1.83 | 1.32 | 72.5 |