Editing Cre 751708 761708 flp QC 2014 11 19-21

11/17/2014: transfect 293 cells in 24 well plate with 2181 (for cre) and 2013(for flp)<br> 

11/18/2014: add recombinase 

freeze-dried cre incubated at 37oC for 3 days and put in RT for two days<br> 

{| width="1147" cellspacing="1" cellpadding="1" border="1"
|-
| 
Flp negative control 

&nbsp;2013<br> 

| 
Flp positive control&nbsp;<br> 

75ul original Flp VLP<br> 

| 
2uM Flp <br> 

from cell culture medium 

| &nbsp; 5uM Flp 
from cell culture medium<br> 

| 2uM Flp 
from 293 cell <br> 

| 5uM Flp 
from 293 cell <br> 

|-
| &nbsp;CRE negative control 
2181<br> 

| CRE positive control 
2.5uM CRE <br> 

(lot#56-375)<br> 

| 
2.5uM<br> 

1st time freeze-dried cre 

(0%Trehalose,5%glycerol)<br> 

| 
2.5uM<br> 

1st time freeze-dried cre 

(5%Trehalose,5%glycerol)<br> 

| 
2.5uM<br> 

1st time freeze-dried cre 

(10%Trehalose,5%glycerol)<br> 

| 
2.5uM<br> 1st time freeze-dried cre 

(15%Trehalose,5%glycerol)<br> 

|-
| &nbsp;&nbsp;&nbsp; 2.5uM cre control&nbsp; 4oC<br> 
(0%Trehalose)<br> 

| 
2.5uM<br> 

2nd time freeze-dried cre 

(0%Trehalose)<br> 

| 
2.5uM<br> 

2nd time freeze-dried cre 

(5%Trehalose)<br> 

| 
2.5uM<br> 

2nd time freeze-dried cre 

(10%Trehalose)<br> 

| 
2.5uM<br> 

2nd time freeze-dried cre 

(15%Trehalose)<br> 

| CRE negative control 
2181<br> 

|-
| &nbsp; 1uM CRE 
(lot#75-1708)<br> 

| 2.5uM CRE 
(lot#75-1708)<br> 

| 5uM CRE 
(lot#75-1708)<br> 

| 1uM CRE 
(lot#76-1708)<br> 

| 2.5uM CRE 
(lot#76-1708)<br> 

| &nbsp;5uM CRE 
(lot#76-1708) 

|}

<br>

11/19/2014: pictures under microscope 24HOURS CELL ARE IN GOOD CODITION 

[[Image:751708 761708 flp QC 2014 11 19.JPG]]<br> 

11/20/2014&nbsp; 48HOURS<br> 

<br> 

{| width="1147" cellspacing="1" cellpadding="1" border="1"
|-
| 
Flp negative control 

&nbsp;2013<br> 

| 
Flp positive control&nbsp;<br> 

75ul original Flp VLP<br> 

| 
2uM Flp <br> 

from cell culture medium 

CELL DIED<br> 

| &nbsp; 5uM Flp 
from cell culture medium<br> 

CELL DIED<br> 

| 2uM Flp 
from 293 cell <br> 

CELL DIED<br> 

| 5uM Flp 
from 293 cell <br> 

CELL DIED<br> 

|-
| &nbsp;CRE negative control 
2181<br> 

| CRE positive control 
2.5uM CRE <br> 

(lot#56-375)<br> 

| 
2.5uM<br> 

1st time freeze-dried cre 

(0%Trehalose,5%glycerol)<br> 

| 
2.5uM<br> 

1st time freeze-dried cre 

(5%Trehalose,5%glycerol)<br> 

| 
2.5uM<br> 

1st time freeze-dried cre 

(10%Trehalose,5%glycerol)<br> 

| 
2.5uM<br> 1st time freeze-dried cre 

(15%Trehalose,5%glycerol)<br> 

|-
| &nbsp;&nbsp;&nbsp; 2.5uM cre control&nbsp; 4oC<br> 
(0%Trehalose)<br> 

| 
2.5uM<br> 

2nd time freeze-dried cre 

(0%Trehalose)<br> 

| 
2.5uM<br> 

2nd time freeze-dried cre 

(5%Trehalose)<br> 

| 
2.5uM<br> 

2nd time freeze-dried cre 

(10%Trehalose)<br> 

| 
2.5uM<br> 

2nd time freeze-dried cre 

(15%Trehalose)<br> 

| CRE negative control 
2181<br> 

|-
| &nbsp; 1uM CRE 
(lot#75-1708)<br> 

| 2.5uM CRE 
(lot#75-1708)<br> 

| 5uM CRE 
(lot#75-1708)<br> 

CELLS ARE NOT IN GOOD CONDITION<br> 

| 1uM CRE 
(lot#76-1708)<br> 

| 2.5uM CRE 
(lot#76-1708)<br> 

| &nbsp;5uM CRE 
(lot#76-1708) 

CELLS ARE NOT IN GOOD CONDITION<br> 

|}

[[Image:751708 761708 flp QC 2014 11 20-C.JPG]]<br> <br> 

11/21/2014 72HOURS, most of the cells are not in good condition<br>

[[Image:751708 761708 flp QC 2014 11 21.JPG]]

[[Category:QC]]