Multiple clones

Subcloning of egfp, rfp, mnatue from pet 28b to pLenti and paav vector

Purpose: Subcloning of egfp, rfp, mnatetune, ifp from pet 28b to pLenti and paav vector .

Date: 2011-8-18

Notebook: Haiyan

Page:003025

Clone ID: pLenti-IFP (226), pLenti-RFP (221),plenti-EGFP (344), plenti -mnatetuen (345), pLenti-BFP (223), paav-mnatetuen (346), pAAV-CMV-IFP (285),pAAV-rfp (347), paav-egfp (348)

Experimenters: Haiyan

Results

Subcloning

Date: 2011-8-18

Digest fragments from pet 28b vector using bamhi+xhoi

Digest paav-mcs or plentiplus with bamhi+xhoi

cut out insert and vector, ligate and transform STBL3

Colony Screening

Date: 2011-8-18

Make miniprep and digest with bamhi+xhoi

Lane 1-4: pLenti-IFP (226) clone 1-4, bamhi+xhoi

Lane 5-8: plenti -mnatetuen (345), clone 1-4, bamhi+xhoi

Lane 9-12: pLenti-RFP (221), clone 1-4, bamhi+xhoi

Lane 13-16: pLenti-BFP (223), clone 1-4, bamhi+xhoi

Lane 2-5: plenti-EGFP (344) clone 1-4, bamhi+xhoi

Lane 6-9: paav-mnatetuen (346), clone 1-4, bamhi+xhoi

Lane 10-11: pAAV-CMV-IFP (285), clone 1-2, bamhi+xhoi

Lane 1-2: pAAV-CMV-IFP (285), clone 3-4, bamhi+xhoi

Lane 3-6: pAAV-rfp (347), clone 1-4, bamhi+xhoi

Lane 7-10: paav-egfp (348), clone 1-4, bamhi+xhoi

Result intepretation/Summary

Got all the positive colonies for above clones

Conclusion

User:Haiyan 08:23, 18 Aug 2011 (EST)