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Duke University/9 June 2009
From 2009.igem.org
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(New page: Measured concentration of DNA fragments <br> Did PCR with fragments 1 and 3 as templates) |
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| - | Measured concentration of DNA fragments | + | [[ Team:Duke | Return ]] |
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| + | ==June 9, 2009== | ||
| + | Gel Extraction | ||
| + | #Weigh cut gel piece(s). Add at least 1 ul binding buffer per mg of gel. | ||
| + | #Place tube at 50-60°C for 10-15 min or until gel is completely dissolved. Vortex every few minutes. | ||
| + | #Add solution to spin column. Centrifuge at max speed for 30s. Discard filtrate. | ||
| + | #Add 300 ul binding buffer to column. Centrifuge for 30s. Discard filtrate. | ||
| + | #Add 500 ul wash buffer. Centrifuge for 30s. Discard filtrate. | ||
| + | #Repeat step 5. | ||
| + | #Centrifuge tubes for at least 2 min. Discard collection tube. | ||
| + | #Obtain 1.7 ml centrifuge tube. Add 40-50 ul elution buffer to spin column. Centrifuge for 1 min. Discard spin column. | ||
| + | |||
| + | *E: Measured concentration of DNA fragments | ||
| + | *R: Very low concentrations of fragments 1 and 3 | ||
| + | *C: Do PCR with fragments as template, greater volume to increase amount | ||
| + | *E: PCR with fragments 1 and 3 as templates | ||
Latest revision as of 16:35, 21 October 2009
June 9, 2009
Gel Extraction
- Weigh cut gel piece(s). Add at least 1 ul binding buffer per mg of gel.
- Place tube at 50-60°C for 10-15 min or until gel is completely dissolved. Vortex every few minutes.
- Add solution to spin column. Centrifuge at max speed for 30s. Discard filtrate.
- Add 300 ul binding buffer to column. Centrifuge for 30s. Discard filtrate.
- Add 500 ul wash buffer. Centrifuge for 30s. Discard filtrate.
- Repeat step 5.
- Centrifuge tubes for at least 2 min. Discard collection tube.
- Obtain 1.7 ml centrifuge tube. Add 40-50 ul elution buffer to spin column. Centrifuge for 1 min. Discard spin column.
- E: Measured concentration of DNA fragments
- R: Very low concentrations of fragments 1 and 3
- C: Do PCR with fragments as template, greater volume to increase amount
- E: PCR with fragments 1 and 3 as templates
