Paul.xu at 19:30, 3 April 2011

2011-04-03T19:30:32Z

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PHP DNA binding experiments 

1:10 dilutioin: take 5 ul from each reaction, add 45 ul water

make 1:5, 1:10, 1:50 and 1:100 dilutions from above 1:10 dilution

add 0.5 ug DNA, 1 ul GRGreen, mix and take a picture

 

[[Image:PHP a b DNA binding.JPG]] 

[[Image:PHP c d DNA binding.JPG]] 

[[Image:PHP e f DNA binding.JPG]] 

--[[User:Paul.xu|Paul.xu]] 16:01, 23 March 2011 (EST) 

[[Image:Compare PHP CBA PEI18 transfection d2.JPG]]

[[Image:Compare PHP CBA PEI18 transfection d3.JPG]]

[[Image:Compare PHP CBA PEI18 transfection d4.JPG]] [[Image:Compare PHP CBA PEI18 transfection 4 3 11.JPG.JPG]]

Transfection:

 

take 5 ul add 45 ul H2O-->1:10 stock

make 1:5, 1:10, 1:50 dilution from 1:10 stock

use 0.5 ug EGFP DNA, 3 ul diluted transfection reagents

[[Image:PHP transfection dilution 14.JPG]]

binding: 1ugDNA, 1ul dye, 3 ul 1:50 (1:5 dilution from 1:10 stock)

Day 2 after transfection --[[User:Paul.xu|Paul.xu]] 10:35, 26 March 2011 (EST)

[[Image:Compare PHP CBA PEI18 transfection 10 folds dilution d1.JPG]]

[[Image:Compare PHP CBA PEI18 transfection exp2 d1]]

 

[[Image:Compare PHP CBA PEI18 transfection exp2 d1.JPG]]

Day1 post transfection

[[Image:Compare PHP CBA PEI18 transfection exp2 d2.JPG]]

day3 [[Image:Compare PHP CBA PEI18 transfection exp2 d5.JPG]]

day4 [[Image:Compare PHP CBA PEI18 transfection exp2 d4.JPG]]

day5 [[Image:Compare PHP CBA PEI18 transfection exp2 4 3 11.JPG.JPG]] 

 

Transfection:

1:10 dilution, 3 ul (or 1 ul), 0.5 ug pCMV-GFP DNA.

GE: GeneExpresso 

PE: PolyExpress 

TR0: without CBA conjugation 

{| cellspacing="1" cellpadding="1" border="1" width="200"
|-
| Sample 
| Code 
|-
| A 
| TR1 
|-
| B 
| TR2 
|-
| C 
| TR3 
|-
| D 
| TR4 
|-
| E 
| TR5 
|-
| F 
| TR6 
|}

 

Dilution of TR3

[[Image:New transfection dilution1.jpg]]

[[Category:Transfection]]

New transfection reagent 2 - Revision history

Paul.xu at 19:30, 3 April 2011