Editing Baculovirus Cre FLP QC 2014 10 09

VLP from sf9 cells. 

Purified FLP: Concentrated from 1L to to 200ml (FLP) by ultrafiltration and purified by gel filtration 

Original FLP: VLP in sf9 cell culture medium<br> 

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

Reporter: 2013 for Flp, 2181 for Cre<br> 

<br> 

Oct. 7 2014 17:00 pm&nbsp;: Transfect 293 cell (70--80%) using lipofectamin 2000<br> 

Oct.8 2014 9:00 am: 20%-30% cells&nbsp; turns red , cells are in good condition, add Flp and&nbsp; Cre to reporter cells<br> 

{| width="822" cellspacing="1" cellpadding="1" border="1"
|-
| 
A1: <br> 

25ul Purified FLP 

| 
A2:<br> 

50ul Purified FLP 

| 
A3:<br> 

75ul Purified FLP 

| 
A4:<br> 

150ul Purified FLP 

| 
A5:<br> 

75ul Original FLP 

| 
A6:<br> 

150ul Original FLP 

|-
| 
B1: 

25ul Purified Cre<br> 

| 
B2: 

50ul Purified Cre<br> 

| 
B3: 

75ul Purified Cre<br> 

| 
B4: 

100ul Purified Cre<br> 

| 
&nbsp;B5: 

150ul Purified Cre<br> 

| 
B6: 

200ul Purified Cre<br> 

|}

Oct.9 2014 9:00 am take photos 

B4,B5 cell are not vey good 

B6 some cells dies, bad<br> [[Image:CRE FLP 239 LIPO2000.JPG]]<br> 

<br> 

Oct. 7 2014 17:00 pm&nbsp;: Split reporter cells into 24well plate, (2013 from another 24 well plate, 2181 from 10cm plate, transfected on Oct,3)<br> 

Oct.8 2014 9:00 am: Flp reporter cells are red ,but in bad condition, only few Cre reporter cells are red 

add Flp and Cre to reporter cells<br> 

{| width="822" cellspacing="1" cellpadding="1" border="1"
|-
| 
A1: <br> 

0ul Purified FLP 

| 
A2:<br> 

25ul Purified FLP 

| 
A3:<br> 

50ul Purified FLP 

| 
A4:<br> 

75ul Purified FLP 

| 
A5:<br> 

100ul Original FLP 

| 
A6:<br> 

150ul Original FLP 

|-
| B1: 
200ul Purified FLP <br> 

| B2: 
50ul Original FLP <br> 

| B3: 
75ul Original FLP<br> 

| B4: 
100ul Original FLP<br> 

| B5: 
100ul Original FLP<br> 

| B6: 
150ul FIRST FILTERED FLP<br> 

|-
| 
C1: 

10ul Purified Cre<br> 

| C2: 
25ul Purified Cre<br> 

| C3: 
50ul Purified Cre<br> 

| C4: 
75ul Purified Cre<br> 

| C5: 
100ul Purified Cre<br> 

| C6: 
125ul Purified Cre<br> 

|-
| D1: 
150ul Purified Cre<br> 

| D2: 
200ul Purified Cre<br> 

| CONTROL<br> 
| <br> 
| <br> 
| <br>
|}

<br> <br> 

[[Image:CRE FLP OLD239.JPG]] 

[[Category:QC]]