Wayne:Laboratory Protocols: Difference between revisions
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primers<br> | primers<br> | ||
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Ethanol precipitation of DNA: Primarily used to clean-up DNA that has been stored in phenol<br> | |||
1. In the 1.5mL epi tube containing the DNA sample to clean, add 1mL of COLD 100% ethanol and 100ul sodium acetate (NaOAc) pH 5.2. Mix by inverting<br> | 1. In the 1.5mL epi tube containing the DNA sample to clean, add 1mL of COLD 100% ethanol and 100ul sodium acetate (NaOAc) pH 5.2. Mix by inverting<br> | ||
2. Place in -20°C freezer for at least 3 hours, overnight is fine<br> | 2. Place in -20°C freezer for at least 3 hours, overnight is fine<br> | ||
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6. Discardsupernatant<br> | 6. Discardsupernatant<br> | ||
7. DrypelletinSpeedVacforupto10minatlowormediumheat<br> | 7. DrypelletinSpeedVacforupto10minatlowormediumheat<br> | ||
8. Re-suspendpelletin1xTEorQiagenAEbuffer | 8. Re-suspendpelletin1xTEorQiagenAEbuffer. Generally use 50-200ul, depending onamountofDNApresent b. ThermomixerisprobablyidealatRT<br> | ||
Revision as of 15:27, 31 January 2013
Put protocols here for:
msats
lab safety
sequencing
pcr
primers
Ethanol precipitation of DNA: Primarily used to clean-up DNA that has been stored in phenol
1. In the 1.5mL epi tube containing the DNA sample to clean, add 1mL of COLD 100% ethanol and 100ul sodium acetate (NaOAc) pH 5.2. Mix by inverting
2. Place in -20°C freezer for at least 3 hours, overnight is fine
3. Spin at 14,000 rpm in a chilled centrifuge at 4°C for 10-15 min
4. Discard supernatant. Careful not to disturb pellet! Pellet may be invisible
5. Re-suspendin1mlfresh70%ethanol,centrifugefor5minat14,000rpm
6. Discardsupernatant
7. DrypelletinSpeedVacforupto10minatlowormediumheat
8. Re-suspendpelletin1xTEorQiagenAEbuffer. Generally use 50-200ul, depending onamountofDNApresent b. ThermomixerisprobablyidealatRT
