Team:TUDelft/Notebook
Notebook
General
Week 24
Week 25
LAB TRAINING
Week 26
Week 27
2/7
Iris and Maartje:
Ampicillin stock
To obtain a 100 mg/ml Ampicillin stock, 500 mg Ampicillin was dissolved in 5 ml milliQ water and filter sterilized using a 45µm filter.
Week 28
8/7
GFP:
- Colonies grew on plate o/n.
- 10ml cultures with one colony picked from the plate [10ml cultures]
- Control (-) by inoculating with a sterile tip LB (no Ab).
9/7
GFP:
- Colonies grew on plate o/n.
- 10ml cultures with one colony picked from the plate [10ml cultures]
- Control (-) by inoculating with a sterile tip LB (no Ab).
13/7
Alicia
Plates of LB agar containing either Chloramphenicol or Ampicillin were prepared following the [agar plate protocol].
14/7
GFP:
2 x 10ml cultures [Liquid starter culture] by inoculating from the cultures stored in the fridge.
15/7
GFP:
- Cultures grew sucessfully.
- 3 minipreps per each culture [Plasmid DNA isolation].
Week 29
Week 30
Week 31
27/7
GFP:
Cultures grew successfully, no growth was seen in negative controls, indicating a successful manipulation.
28/7
GFP:
Cultures grew successfully, no growth was seen in negative controls, indicating a successful manipulation.
3x Minipreps [Plasmid DNA isolation] were done, resulting in ratios of absorbance A260/A280 below 1.8, which indicates protein contamination.
29/7
GFP:
2x 10ml cultures of GFP from colony 1 [Liquid starter culture] were cultured again overnight to ensure good quality stock of plasmids without contamination.
30/7
GFP:
2 x 10ml cultures [Liquid starter culture] by inoculating from the cultures stored in the fridge.
31/7
GFP:
Phage insert was prepared to be transformed into BL21 for phage engineering. PCR using KOD Xtreme [KOD PCR] was used to amplify with primers GFP_1.1_Fw & Rv, GFP_0.6_Fw & Rv, GFP_4.3_Fw & Rv using eGFP plasmid as a template from minipreps made yesterday. The 50 µl PCR products were loaded into a 1% agarose gel with Sybr [AGAROSE GEL].
Results indicate the expected band size amplification. A gel-purification was performed to escise the bands obtaining 85 ng/µl (GFP_insert_0.6), 110 ng/µl (GFP_insert_1.1), 93 ng/µl (GFP_insert_4.3) [GEL PURIFICATION].
Week 32
Week 33
Week 34
Week 35
Week 36
4/9
Alicia:
Plates of LB agar containing either Chloramphenicol, kanamycin or Ampicillin were prepared following the [agar plate protocol].
Week 38
Week 39
Week 40
Week 41
Week 42