Difference between revisions of "Team:UCLA/Notebook/Honeybee Silk/4 May 2015"
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#Pre heat plates and SOC at 37 C | #Pre heat plates and SOC at 37 C | ||
#Thaw 2 aliquots of 50 uL of competent cells on ice for 10 minutes | #Thaw 2 aliquots of 50 uL of competent cells on ice for 10 minutes | ||
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#Add 2 ul of non-purified ligated product | #Add 2 ul of non-purified ligated product | ||
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#Incubate for 10 minutes on ice | #Incubate for 10 minutes on ice | ||
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#Rescue in 200 uL SOC at 37°C for ~1 hour | #Rescue in 200 uL SOC at 37°C for ~1 hour | ||
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#Plate 100 uL in 1:1 and 1:10 ratios onto chloramphenicol plates | #Plate 100 uL in 1:1 and 1:10 ratios onto chloramphenicol plates | ||
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#Spread with beads | #Spread with beads | ||
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#Incubate plates at 37 C (put in at 3:30) | #Incubate plates at 37 C (put in at 3:30) | ||
Latest revision as of 22:09, 29 May 2015
5/4 Transformation of Silk and Prom + Silk
- Using Sri’s chemically competent cells
- Using ligation product from 4/30 (not purifying the ligation product as per Eric’s recommendation)
- Pre heat plates and SOC at 37 C
- Thaw 2 aliquots of 50 uL of competent cells on ice for 10 minutes
- Add 2 ul of non-purified ligated product
- Incubate for 10 minutes on ice
- Rescue in 200 uL SOC at 37°C for ~1 hour
- Plate 100 uL in 1:1 and 1:10 ratios onto chloramphenicol plates
- Spread with beads
- Incubate plates at 37 C (put in at 3:30)