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:
- 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
|