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Engenharia genética/Microscopia de fluorescência e SDS-PAGE: diferenças entre revisões

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MPCF (discussão | contribs)
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MPCF (discussão | contribs)
187 edições
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Linha 6:
 
 
1.# Spin your sample for 10-20s
2.# Prepare two eppendorf tubes with 20 uL SDS-sample buffer
2.# Add 20 uL of the samples to the tubes with the SDS-sample buffer, do not transfer cell debris!
3.# Boil the tubes for 5 min.
4.# Load 10 uL of the mixture on the SDS-PAGE.
5.# Run the gel for 30 min – 1 hour at 200v
 
While the gel is running do the following:
7.# Prepare 3 eppendorf tubes with 1 mL water in each. ▼
 
8.# Pick three colonies from the plate marked (+) and resuspend each colony in one of the tubes.▼
▲7. Prepare 3 eppendorf tubes with 1 mL water in each.
# Put 10 uL from each tube on a microscopic slide.
▲8. Pick three colonies from the plate marked (+) and resuspend each colony in one of the tubes.
9.# Put 10a uLcoverslip fromon eachtop tube on aof microscopicthe slideliquid.
11.# Observe the cells using fluorescent microscopy▼
10. Put a coverslip on top of the liquid.
12.# Design the primers that was used for the amplification of GFP with the addition of the sequences needed for gap-repair▼
▲11. Observe the cells using fluorescent microscopy
 
▲12. Design the primers that was used for the amplification of GFP with the addition of the sequences needed for gap-repair
 
 
Obtido em "https://pt.wikibooks.org/wiki/Engenharia_genética/Microscopia_de_fluorescência_e_SDS-PAGE"