Difference between revisions of "SDS Page-Gel for all the bovine casein inserts"
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Thus we have to add total 120ul of dye | Thus we have to add total 120ul of dye | ||
Boil all the tubes for 10 mins at 95C ( | Boil all the tubes for 10 mins at 95C (in order to denature the protein structure to linear) | ||
'''Results''' | |||
'''Gel#2''' | |||
[[File:Gel2 17Jan2015 labeled.jpg]] | |||
Revision as of 01:00, 20 January 2015
17 January 2015: SDS Page-Gel All 5 Bovine Casein Inserts
Location: BioCurious
Participants: Lafia, Johan, Meenakshi, Emi, Rachel
Notes: Emi
Aims: Confirmation of bovine casein inserts from liquid culture of yeast transformed cells
Materials
Gel loading tips
Tris-Acelate SDS Running Buffer novex life tech
Bio-Rad pipette tips
Native gels... (need details)
ladder 20x
Casein
Cytosin
Transformed negative control
Bov alpha s1 (1) Transformed liqid culture
Bov beta B (8) Transformed liquid culture
Bov alpha S2 (3) Transformed liquid culture
Bov beta A2 (6B) Transformed liquid culture
Bov kappa (7F) Transformed liquid culture
Chymosin + casein (Chymosin cuts casein and separates it to 2 bands)
Ladder
Protocol
Centrifuge the falcon tubes, 5 minutes 2800xg
Take 500ul of distilled water in eppendorf tube + 1 tiny drop of casein (use 200ul hp take it) - 2times
Take 15ul of chymosin +1 tiny drop of casein in 250ul of water
Take 250ul of 7F (Bov Kappa) + 15ul of Chymosinin Take 250ul of water and 15ul of chymosin
Take 40ul of 1, 8, 3B, 6B, +7F also the negative control to eppendorf tubes
Preparation fo 20x running buffer
Pour 50ul of 10x running buffer; make it to 450ul by adding the distilled water - total 500ul
20ul of loading dye to each tubes except casein; chymosin and casein; chymosin and 7F
Thus we have to add total 120ul of dye
Boil all the tubes for 10 mins at 95C (in order to denature the protein structure to linear)
Results
Gel#2
