Baculovirus Cre FLP QC 20141009 20141013

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VLP from sf9 cells.

Final Purified FLP: Concentrated from 1L to to 200ml (FLP) by ultrafiltration, purified by gel filtration, then ultrafiltrated again, filtered on 10/10/2014

Original FLP: VLP in sf9 cell culture medium

Purified Cre: Concentrated from 1L to to 200ml (FLP) by ultrafiltration , dialyzed , purified by gel filtration , filtered on 10/08/2014

Final Purified Cre: Concentrated from 1L to to 200ml (FLP) by ultrafiltration , dialyzed , purified by gel filtration ,then ultrafiltrated again, filtered on 10/10/2014

Reporter: 2013 for Flp, 2181 for Cre


Oct. 9 2014 10:00 am : split 293 cells

Oct. 9 2014 18:00 am Transfect 293 cell (70--80%) using lipofectamin 2000

Oct.10 2014 10:00 am: 40%-50% cells  turns red , cells are in good condition, add Flp and  Cre to reporter cells

A1:

75ul original FLP

A2:

0 ul Purified FLP

A3:

10ul Final Purified FLP

A4:

25ul Final Purified FLP

A5:

50ul Final Purified FLP

A6:

75ul Final Purified FLP

B1:

100ul Final Purified FLP

 B2:

150ul Final Purified FLP

 B3:

200ul Final Purified FLP

 B4:

50ul Final Purified FLP (unfiltered)

 B5:

75ul Final Purified FLP (unfiltered)

 B6:

150ul Final Purified FLP (unfiltered)

C1:

75ul Purified Cre

 C2:

0ul PurifiedCre

 C3:

10ul Final Purified Cre

 C4:

25ul Final Purified Cre

 C5:

50ul Final Purified Cre

 C6:

75ul Final Purified Cre

D1:

100 ul Final Purified Cre

 D2:

150 ul Final Purified Cre

D3:

200 ul Final Purified Cre

D4:

50ul Final Purified CRE (unfiltered)

D5:

75ul Final Purified CRE (unfiltered)


D6:

150ul Final Purified CRE (unfiltered)



Oct.12 2014  take photos


picture of 2014/10/16

A1:

75ul original FLP

A2:

0 ul Purified FLP

A3:

10ul Final Purified FLP

A4:

25ul Final Purified FLP

A5:

50ul Final Purified FLP

A6:

75ul Final Purified FLP

B1:

100ul Final Purified FLP

 B2:

150ul Final Purified FLP

 B3:

200ul Final Purified FLP

C1: 75ul Purified Cre

C2: 0ul PurifiedCre C3: 10ul Final Purified Cre C4: 25ul Final Purified Cre C5: 50ul Final Purified Cre
C6: 75ul Final Purified Cre D1:

100 ul Final Purified Cre

 D2:

150 ul Final Purified Cre

D3:

200 ul Final Purified Cre



Unfiltered: all contaminated





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