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Team:Newcastle/22 June 2010
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The 5ml culture set up yesterday was pelleted down using the microcentrifuge, the pellet was resuspended in buffer P1. | The 5ml culture set up yesterday was pelleted down using the microcentrifuge, the pellet was resuspended in buffer P1. | ||
Lysis buffer P2 was used to release the DNA from the cell (this buffer contains RNAse to help reduce RNA contamination). PE buffer containing ethanol was used as a wash. We lysed the cells for 1 minute, gently inverting the tube. We neutalised the lysis with N3 buffer and centrifuged for 10minutes, only the '''plasmid DNA was left in suspension'''. The DNA was eluted at low salt concentration through a column membrane. | Lysis buffer P2 was used to release the DNA from the cell (this buffer contains RNAse to help reduce RNA contamination). PE buffer containing ethanol was used as a wash. We lysed the cells for 1 minute, gently inverting the tube. We neutalised the lysis with N3 buffer and centrifuged for 10minutes, only the '''plasmid DNA was left in suspension'''. The DNA was eluted at low salt concentration through a column membrane. | ||
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| + | ===Digest=== | ||
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| + | ===Gel extraction=== | ||
===Set up ligation=== | ===Set up ligation=== | ||
Revision as of 10:02, 24 June 2010
Tuesday
Contents |
Qiagen miniprep: Plasmid extraction
The 5ml culture set up yesterday was pelleted down using the microcentrifuge, the pellet was resuspended in buffer P1. Lysis buffer P2 was used to release the DNA from the cell (this buffer contains RNAse to help reduce RNA contamination). PE buffer containing ethanol was used as a wash. We lysed the cells for 1 minute, gently inverting the tube. We neutalised the lysis with N3 buffer and centrifuged for 10minutes, only the plasmid DNA was left in suspension. The DNA was eluted at low salt concentration through a column membrane.
