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Team:Hong Kong HKUST/notebook/mod3
From 2013.igem.org
June 2013
· Inoculation of pBlueScript KS(+) for training
· Autoclave basic materials
· Preparing LB
· LB-Ampicillin plates poured
Dry lab
Protocols review
August 2013
August 5
GFP (25)PCR Check with VR VF2
- Gel check(overan)
BBa_J176171
- Digestion using SpeI and PstI-HF to extract the backbone
PolyA / BBa_K404108
- Digestion using XbaI and PstI-HF
- Gel electrophoresis
- Run Gel
Inoculation of pSB1C3
August 6
GFP (25)
- Repeat PCR check
- Run Gel
pSB1C3
- Plasmid extraction
PolyA / BBa_K404108
- Gel purification
- Gel check
BBa_J176171
- PCR clean up
BBa_K929000 & BBa_J52034
- Transformation using CAM plate
MLS
- PCR amplify MLS in RFC 10 and 25 format
August 7
MLS (RFC 10 & 25)
- PCR clean up
- Nanodrop for the PCR clean up product
- Gel check for the PCR clean up product
- Digestion using EcoRI and PstI
PolyA / BBa_K404108
- Gel check
pSB1C3
- Digest with EcoRI & PstI
- Run gel and cut gel
GFP(25)
- Digestion with EcoRI & SpeI
- Run gel and cut gel
August 8
GFP25 & pSB1C3
- Gel purification
- Nanodrop
- Gel check
polyA
- Digestion with EcoRI and XbaI
- PCR clean up
BBa_K929000 & BBa_J52034
- Inoculation
August 9
BBa_K929000 & BBa_J52034
- Plasmid extraction: J52034 is not transformed
MLS.pSB1C3 & GFP.PolyA
- Ligation
- Transformation to SURE on CAM plate
