ELN - User contributions [en]

File:Transfection reagent comparison 4-14-2011.jpg

2011-04-15T13:49:02Z

Ztao: uploaded a new version of "File:Transfection reagent comparison 4-14-2011.jpg"

PDB HIS Nusa Strep II

2011-04-13T20:56:28Z

Ztao:

from left to right: molecular weight, last one is PDB_HIS_Nusa cut by XhoI and HindIII. Other bands are for another project.

[[Image:PDB_His_Nusa_cut_by_XhoI_and_HindIII_4-13-2011.JPG|187x144px]]--[[User:Ztao|Ztao]] 15:56, 13 April 2011 (EST)

[[Category:Cloning]]

File:PDB His Nusa cut by XhoI and HindIII 4-13-2011.JPG

2011-04-13T20:54:41Z

Ztao:

Taq Purification April

2011-04-13T20:51:58Z

Ztao:

1ul Taq containing culture was use to do the PCR. Total volume of PCR is 25 ul. 

from left to right: molecular marker, positive control, sample.

[[Image:Taq activity 4-13-2011.JPG|162x321px|Taq activity 4-13-2011.JPG]] --[[User:Ztao|Ztao]] 15:51, 13 April 2011 (EST)

[[Category:Protein_Purification]]

File:Taq activity 4-13-2011.JPG

2011-04-13T20:43:07Z

Ztao:

PKA ELISA 4-8-2011

2011-04-08T18:24:15Z

Ztao:

Notebook Zhen Tao #11 

Page 2101~2102 

all samples are diluted 1/400 with blocking solution; Control: only 2nd antibody (anti-human) was used; sample 29 and 447667~447669: prostate cancer people serum; sample 447670~447672: colon cancer people serum; sample 447673~447675: breast cancer people serum; 

sample 31~146 are normal people serum; positive: anti-PKA-C-alpha is used

15 min, instead of 30 min was used here after adding TMB solution. 

{| cellspacing="1" cellpadding="1" border="1" style="width: 752px; height: 555px;"
|-
| samples 
| OD450-1 
| OD450-2 
| OD450-3 
| average of all normal serum 
| average of cancer sample 
| ration of cancer/normal 
|-
| control 
| 0.005 
| 0.006 
| 0.005 
| 
| 
| 
|-
| 447667 
| 0.856 
| 0.841 
| 0.850 
| 
| 0.844 
| 3.706 
|-
| 447668 
| 0.501 
| 0.508 
| 0.526 
| 
| 0.506 
| 2.224 
|-
| 447669 
| 0.323 
| 0.331 
| 0.309 
| 
| 0.316 
| 1.387 
|-
| 447670 
| 0.290 
| 0.289 
| 0.290 
| 
| 0.284 
| 1.249 
|-
| 447671 
| 0.143 
| 0.146 
| 0.152 
| 
| 0.142 
| 0.622 
|-
| 447672 
| 0.468 
| 0.505 
| 0.547 
| 
| 0.501 
| 2.202 
|-
| 447673 
| 0.895 
| 0.545 
| 0.585 
| 
| 0.670 
| 2.942 
|-
| 447674 
| 0.153 
| 0.160 
| 0.169 
| 
| 0.155 
| 0.682 
|-
| 31 
| 0.190 
| 0.193 
| 0.201 
| rowspan="12" | 0.228 
| 
| 
|-
| 32 
| 0.279 
| 0.281 
| 0.318 
| 
| 
|-
| 33 
| 0.232 
| 0.255 
| 0.242 
| 
| 
|-
| 34 
| 0.177 
| 0.168 
| 0.151 
| 
| 
|-
| 35 
| 0.412 
| 0.393 
| 0.418 
| 
| 
|-
| 39 
| 0.205 
| 0.208 
| 0.211 
| 
| 
|-
| 40 
| 0.106 
| 0.124 
| 0.124 
| 
| 
|-
| 132 
| 0.285 
| 0.286 
| 0.260 
| 
| 
|-
| 133 
| 0.167 
| 0.172 
| 0.182 
| 
| 
|-
| 135 
| 0.151 
| 0.153 
| 0.166 
| 
| 
|-
| 136 
| 0.058 
| 0.590 
| 0.156 
| 
| 
|-
| 146 
| 0.313 
| 0.291 
| 0.269 
| 
| 
|-
| positive 
| 1.718 
| 1.824 
| 1.869 
| 
| 
| 
|}

 

 --[[User:Ztao|Ztao]] 13:03, 8 April 2011 (EST) 

 

[[Category:ELISA]]

PKA ELISA 4-7-2011

2011-04-07T18:32:51Z

Ztao:

Notebook Zhen Tao #10 

Page 2097~2098 

 

all samples were diluted 1/2000 with blocking solution; Control: only 2nd antibody (anti-human) was used; sample 31~40: normal people serum; sample 447667~447669: prostate cancer people serum; sample 447670~447672: colon cancer people serum; sample 447673~447674: breast cancer people serum; positive: anti-PKA-C-alpha was used (1/10000)

OD450 data for normal and cancer samples are corrected by substrating the average of control (0.018). 

{| cellspacing="1" cellpadding="1" border="1" style="width: 550px; height: 434px;"
|-
| samples 
| OD450-1 
| OD450-2 
| OD450-3 
| average of normal serum 
| cancer/normal 
|-
| control 
| 0.017 
| 0.018 
| 0.020 
| 0.018 
| rowspan="8" | 
|-
| 31 
| 0.107 
| 0.098 
| 0.109 
| rowspan="7" | 0.124 
|-
| 32 
| 0.169 
| 0.157 
| 0.165 
|-
| 33 
| 0.144 
| 0.133 
| 0.152 
|-
| 34 
| 0.092 
| 0.126 
| 0.071 
|-
| 35 
| 0.154 
| 0.204 
| 0.159 
|-
| 39 
| 0.122 
| 0.130 
| 0.098 
|-
| 40 
| 0.065 
| 0.087 
| 0.077 
|-
| 447667 
| 0.499 
| 0.650 
| 0.463 
| 
| 4.317 
|-
| 447668 
| 0.387 
| 0.492 
| 0.346 
| 
| 3.280 
|-
| 447669 
| 0.170 
| 0.257 
| 0.117 
| 
| 1.455 
|-
| 447670 
| 0.232 
| 0.246 
| 0.188 
| 
| 1.782 
|-
| 447671 
| 0.077 
| 0.082 
| 0.080 
| 
| 0.638 
|-
| 447672 
| 0.405 
| 0.433 
| 0.410 
| 
| 3.342 
|-
| 447673 
| 0.274 
| 0.281 
| 0.258 
| 
| 2.176 
|-
| 447674 
| 0.095 
| 0.095 
| 0.101 
| 
| 0.777 
|-
| postitive 
| 0.964 
| 0.976 
| 0.989 
| 
| 7.847 
|}

 --[[User:Ztao|Ztao]] 13:29, 7 April 2011 (EST)

[[Category:ELISA]]

PKA ELISA 4-6-2011

2011-04-06T19:28:56Z

Ztao:

Notebook Zhen Tao #10 

Page 2096 

All samples are diluted 1/10000 with blocking solution (1%BSA, 10%glycerol in TBS). 

Control: only 2nd antibody (anti-human) was used; sample 31~40 are normal people serum; sample 447667~447669: prostate cancer people serum; sample 447670~447672: colon cancer people serum; sample 447673~447674: breast cancer people serum. 

{| cellspacing="1" cellpadding="1" border="1" style="width: 419px; height: 477px;"
|-
| Samples 
| OD450-1 
| OD450-2 
| OD450-3 
| average 
| average of normal serum sample
|-
|
control 

| 0.019 
| 0.017 
| 0.016 
| 0.017 
| rowspan="8" | 0.036
|-
| 31 
| 0.031 
| 0.033 
| 0.034 
| 0.033 
|-
| 32 
| 0.044 
| 0.043 
| 0.043 
| 0.043 
|-
| 33 
| 0.037 
| 0.037 
| 0.045 
| 0.040 
|-
| 34 
| 0.029 
| 0.035 
| 0.046 
| 0.037 
|-
| 35 
| 0.043 
| 0.042 
| 0.040 
| 0.042 
|-
| 39 
| 0.032 
| 0.033 
| 0.032 
| 0.032 
|-
| 40 
| 0.027 
| 0.024 
| 0.026 
| 0.026 
|-
| 447667 
| 0.108 
| 0.094 
| 0.107 
| 0.103 
|
|-
| 447668 
| 0.087 
| 0.077 
| 0.082 
| 0.082 
|
|-
| 447669 
| 0.039 
| 0.036 
| 0.030 
| 0.035 
|
|-
| 447670 
| 0.051 
| 0.045 
| 0.042 
| 0.046 
|
|-
| 447471 
| 0.030 
| 0.031 
| 0.027 
| 0.029 
|
|-
| 447672 
| 0.035 
| 0.035 
| 0.039 
| 0.036 
|
|-
| 447673 
| 0.058 
| 0.063 
| 0.054 
| 0.058 
|
|-
| 447674 
| 0.023 
| 0.025 
| 0.026 
| 0.025 
|
|-
| positive control 
| 0.345 
| 0.339 
| 0.342 
| 0.342 
|
|}

 --[[User:Ztao|Ztao]] 14:26, 6 April 2011 (EST)

[[Category:ELISA]]

File:4-6-2011 PKA.xls

2011-04-06T19:13:05Z

Ztao:

PKA ELISA 4-5-2011

2011-04-05T20:35:35Z

Ztao:

Notebook zhen tao #10

Page 2092

plate was coated with 2ug/ml PKA-C-alpha in TBS; 100 ul/well; then blocked by 1% BSA in TBS and 10% glycerol. each sample was diluted 1/10000 with blocking buffer.

 

{| width="200" cellspacing="1" cellpadding="1" border="1"
|-
| samples
| OD450-1
| OD450-2
| OD450-3
| average
|-
| control
| 0.025
| 0.021
| 0.019
| 0.022
|-
| 31
| 0.056
| 0.056
| 0.056
| 0.056
|-
| 32
| 0.078
| 0.080
| 0.080
| 0.079
|-
| 447673
| 0.110
| 0.122
| 0.155
| 0.129
|-
| 447674
| 0.054
| 0.059
| 0.056
| 0.056
|-
| positive control
| 0.569
| 0.557
| 0.556
| 0.561
|}
--[[User:Ztao|Ztao]] 15:35, 5 April 2011 (EST)
[[Category:ELISA]]

PKA ELISA 4-1-2011

2011-04-01T20:30:56Z

Ztao: Created page with "Notebook: zhen tao #10 Page: 2085 normal serum #31, cancer serum $447673 and anti-PKA_c alpha are used. {| cellspacing="1" cellpadding="1" border="1" width="..."

Notebook: zhen tao #10 

Page: 2085 

normal serum #31, cancer serum $447673 and anti-PKA_c alpha are used. 

 

{| cellspacing="1" cellpadding="1" border="1" width="200"
|-
| 
| 1 
| 2 
| 3 
| 4 
| 5 
| 6 
| 7 
| 8 
| 9 
| 10 
| 11 
| 12 
|-
| B 
| 0.067 
| 0.071 
| 0.081 
| 0.094 
| 0.109 
| 0.120 
| 0.218 
| 0.230 
| 0.248 
| 0.192 
| 0.209 
| 0.162 
|-
| C 
| 0.277 
| 0.267 
| 0.263 
| 0.627 
| 0.638 
| 0.674 
| 0.353 
| 0.355 
| 0.353 
| 0.080 
| 0.079 
| 0.077 
|}

 B1~B3: #31 1/10000 

B4~B6: #31 1/5000

B7~B9: #31 1/2000

B10~B12: #447673 1/10000

C1~C3: #447673 1/5000

C4~C6: #447673 1/2000

C7~C9: anti-PKA-C alpha 1/10000

C10~C12: control.

2nd antibody: 1/10000
--[[User:Ztao|Ztao]] 15:30, 1 April 2011 (EST)

File:VP16 and PKA 3-30-2011.jpg

2011-04-01T14:01:52Z

Ztao:

Taq Purification

2011-03-29T19:29:43Z

Ztao:

From left to right: Taq protein number 2, 1, positive control and molecular standard. Templete is PEDFR. Total PCR volume is 25 ul.

 

[[Image:Taq activity check.JPG|146x303px|Taq activity check.JPG]] --[[User:Ztao|Ztao]] 08:42, 15 March 2011 (EST)

 

From left to right: molecular marker, positive control, Taq number 3, 4 and 5. Use PEDFR as templete.

[[Image:Taq activity check 2.JPG|179x167px|Taq activity check 2.JPG]] --[[User:Ztao|Ztao]] 14:02, 15 March 2011 (EST)

 

From left to right: molecular weight, positive control, Taq to be purified.

[[Image:Activity of Taq.JPG|144x235px|Activity of Taq.JPG]]

--[[User:Ztao|Ztao]] 12:57, 17 March 2011 (EST)

 

From left to right: molecular weight, positive control, orignal Taq, flow through and wash

[[Image:Activity of Taq 2.JPG|210x190px|Activity of Taq 2.JPG]]

 --[[User:Ztao|Ztao]] 15:41, 18 March 2011 (EST)

From left to right: molecular weight, positive control, sample from big, sample from small, sample from solution

[[Image:Activity_of_Taq_on_3-29-2011.JPG|165x188px]]--[[User:Ztao|Ztao]] 14:29, 29 March 2011 (EST)

[[Category:Protein_Purification]]

File:Activity of Taq on 3-29-2011.JPG

2011-03-29T19:27:52Z

Ztao:

File:PLVX-PLUS2 cut by BamHI and XhoI.JPG

2011-03-29T13:53:22Z

Ztao:

Category:ELISA

2011-03-28T21:56:43Z

Ztao: Blanked the page

PKA ELISA dev 1

2011-03-28T21:56:16Z

Ztao: Created page with "{| cellspacing="1" cellpadding="1" border="1" style="width: 360px; height: 282px;" |- | sample | OD450-1 | OD450-2 | OD450-3 | Average |- | 31 | 0.118 | 0.128 | 0.125 | ..."

{| cellspacing="1" cellpadding="1" border="1" style="width: 360px; height: 282px;"
|-
|
sample

| OD450-1
| OD450-2
| OD450-3
| Average
|-
| 31
| 0.118
| 0.128
| 0.125
| 0.124
|-
| 32
| 0.147
| 0.151
| 0.147
| 0.148
|-
| 33
| 0.125
| 0.129
| 0.123
| 0.126
|-
| 34
| 0.123
| 0.130
| 0.134
| 0.129
|-
| 447673
| 0.203
| 0.226
| 0.237
| 0.222
|-
| 447674
| 0.122
| 0.125
| 0.126
| 0.124
|-
| 447675
| 0.151
| 0.159
| 0.158
| 0.156
|-
| positive control
| 0.398
| 0.408
| 0.381
| 0.396
|-
| control
| 0.089
| 0.099
| 0.097
| 0.095
|}

Plate is coated with PKA-C alpha with OD280 = 0.075;

Blocked by using 1%BSA, 10% glycerol in TBS;

Serum or 1st antibody is diluted 10,000 times with blocking buffer;

smaples 31~34 are serum from normal people; samples 447673~75 are from breast cancer people; positive control is anti-PKA-C alpha; control: no serum or 1st antibody was used, only 2nd antibody is used. 

 --[[User:Ztao|Ztao]] 14:19, 25 March 2011 (EST)

 

 

 

 

[[Category:ELISA]]

PLVXPlus2-IRES-mCherry

2011-03-25T20:09:19Z

Ztao:

pLVX_Plus-IRES-mCherry is cut by BamHI and XhoI.

[[Image:PLVXPlus2-IRES-mCherry BamHI XhoI.JPG]]

--[[User:Ztao|Ztao]] 13:56, 15 March 2011 (EST)

 

Clones 1 to 9 analyzed by EcoRI and KnpI. From left to right: molecular weight, clones 1 to 9.

[[Image:Clones 1 to 10 cut by ECorI and KpnI.JPG]] --[[User:Ztao|Ztao]] 12:50, 17 March 2011 (EST)

 

from left to right: molecular weight, next two lines is for another project, line 4 and 5 are clone 1 and 2.

[[Image:Clones 1 and 2 cut by NdeI and EcoRI.JPG|137x160px|Clones 1 and 2 cut by NdeI and EcoRI.JPG]]

--[[User:Ztao|Ztao]] 08:48, 18 March 2011 (EST)

 

Clones 1 to 10 (minipreped on 3/24/2011) analyzed by EcoRI and KnpI. From left to right: molecular weight, clones 1 to 10. 

[[Image:ECroRI_and_KpnI_cut.JPG]] --[[User:Ztao|Ztao]] 15:09, 25 March 2011 (EST)

[[Category:Cloning]]

File:ECroRI and KpnI cut.JPG

2011-03-25T20:07:32Z

Ztao:

3-25-2011 RT-PCR

2011-03-25T19:52:21Z

Ztao:

Notebook: ZHEN TAO #10

Page: 002072

2ul sample 

1ul WPRE-F primer 

1ul WPRE-R primer 

12.5 ul Mastermix (Roche) 

8.5 ul H2O 

Total 25 ul. 

 

1) 50C 2 min; 

2) 95C 10 min; 

3) 95C 15 sec; 

4) 60C 1 min; 

5) repeat step 3 and 4 for 40 times 

 

{| width="200" cellspacing="1" cellpadding="1" border="1"
|-
| tube number 
| 1 
| 2 
| 3 
| 4 
| 5 
| 6 
| 7 
|-
| position in plate 
| D3 
| D4 
| D5 
| D6 
| D7 
| D8 
| D9 
|-
| sample 
| #13 
| #14 
| #13 
| #14 
| #13 
| #14 
| H2O 
|-
| treatment 
| colspan="2" | untreated
| colspan="2" | lysis and then DNAse
| colspan="2" | DNAse and then lysis
| 
|-
| CT value 
| 15.7 
| 15.7 
| 24.6 
| 25.3 
| 24.6 
| 23.3 
| 28.3 
|}

 

Treatment: 

Untreated: 2ul virus mix with 18 ul TBS, and use 2 ul to do RT-PCR;

Lysis and then DNAse: 12 ul virus mix with 108 ul lysis buffer; RT, 10 min; add 2 ul 1M Tris pH8.0; add 1ul DNAse; RT, 30 min; 75C 10 min; use 2 ul to do RT-PCR;

DNAse then lysis: in 12 ul virus, add 1 ul DNAse, RT 30 min; 75C 10 min; add 108 ul lysis buffer; RT 10 min; use 2 ul to do RT-PCR. 

 --[[User:Ztao|Ztao]] 14:48, 25 March 2011 (EST)

 

 

 

[[Category:QPCR_RT-PCR]]

File:Activity of Taq 2.JPG

2011-03-18T20:39:05Z

Ztao:

File:Clones 1 and 2 cut by NdeI and EcoRI.JPG

2011-03-18T13:46:36Z

Ztao:

File:Activity of Taq.JPG

2011-03-17T17:55:15Z

Ztao:

File:Clones 1 to 10 cut by ECorI and KpnI.JPG

2011-03-17T17:47:40Z

Ztao:

File:Taq activity check 2.JPG

2011-03-15T18:59:59Z

Ztao:

File:PLVXPlus2-IRES-mCherry BamHI XhoI.JPG

2011-03-15T18:55:04Z

Ztao:

File:Taq activity check.JPG

2011-03-15T13:38:09Z

Ztao:

Transfection reagents comparison 4

2011-02-22T14:19:40Z

Ztao:

Notebook: Zhen Tao #100

Page: 001954

New transfection reagent: 3, 6 and 9 ul/well

PEIMAX: 1, 3 and 5 ul/well

Turbo: 1, 3 and 5 ul/well

Use 50 ul DMEM(-) to prepare each transfection reagent.

Plasmid: 216390 (0.74 ug/ul); 1ug/well; prepared in 50 ul DMEM(-).

Packaging Mix: v:g=1:3 (0.1ug/ul); 3ul/well; add to 50 ul DMEM(-) containing plasmid 216390.

[[Image:Transfection reagent comparison.JPG]]

3 day after transfection. --[[User:Ztao|Ztao]] 11:00, 21 February 2011 (EST)

[[Image:Transfection_reagent_comparison_2_day_after_transfection.JPG]]

4 day after transfection. --[[User:Ztao|Ztao]] 09:19, 22 February 2011 (EST)

[[Category:Transfection]]

File:Transfection reagent comparison 2 day after transfection.JPG

2011-02-22T14:18:26Z

Ztao:

Subramanian 1 shRNA-5

2011-02-22T14:18:05Z

Ztao:

Notebook: ZHEN TAO #100 Page:001950

In each well, which contains 1 ml cell medium, add 25 or 50 ul virus pre-prepared in 0.5 ml DMEM(+). 

[[Image:Subramanian 1 shRNA 5 comparison of different lentivirus day 1.JPG]]

1 day after transfection. --[[User:Ztao|Ztao]] 09:20, 18 February 2011 (EST)

 

[[Image:Subramanian 1 shRNA 5 comparison of different lentivirus 4 day after transfection.JPG]]

4 day after transfection --[[User:Ztao|Ztao]] 10:52, 21 February 2011 (EST)

[[Image:Subramanian_1_shRNA_5_comparison_of_different_lentivirus_5_day_after_transfection.JPG]]

5 day after transfection. --[[User:Ztao|Ztao]] 09:18, 22 February 2011 (EST)

[[Category:Lentivirus]]

File:Subramanian 1 shRNA 5 comparison of different lentivirus 5 day after transfection.JPG

2011-02-22T14:16:38Z

Ztao:

Subramanian 1 shRNA-4

2011-02-22T14:16:08Z

Ztao:

notebook: ZHEN TAO #100

Page:

1, 5 or 10 ul lentivirus (192, 390, 881 and 882) were added to 12 well plate

[[Image:Subramanian 1 shRNA 4 comparison of different lentivirus.JPG]]

1 day after transfection --[[User:Ztao|Ztao]] 10:54, 17 February 2011 (EST)

 

[[Image:Subramanian 1 shRNA 4 comparison of different lentivirus day 2.JPG]]

2 day after transfection --[[User:Ztao|Ztao]] 09:11, 18 February 2011 (EST)

 

[[Image:Subramanian 1 shRNA 4 comparison of different lentivirus 5 day after transfection.JPG]]

5 day after transfection --[[User:Ztao|Ztao]] 10:50, 21 February 2011 (EST)

[[Image:Subramanian_1_shRNA_4_comparison_of_different_lentivirus_6_day_after_transfection.JPG]]

6 day after transfection. --[[User:Ztao|Ztao]] 09:16, 22 February 2011 (EST)

[[Category:Lentivirus]]

File:Subramanian 1 shRNA 4 comparison of different lentivirus 6 day after transfection.JPG

2011-02-22T14:14:23Z

Ztao:

File:Transfection reagent comparison.JPG

2011-02-21T15:55:39Z

Ztao:

File:Subramanian 1 shRNA 5 comparison of different lentivirus 4 day after transfection.JPG

2011-02-21T15:50:56Z

Ztao:

File:Subramanian 1 shRNA 4 comparison of different lentivirus 5 day after transfection.JPG

2011-02-21T15:49:03Z

Ztao:

File:New Transfection Reagent Test.JPG

2011-02-18T17:07:07Z

Ztao:

Lentivirus pLVX-PEDFR-1

2011-02-18T14:25:01Z

Ztao:

Notebook: ZHEN TAO #100

Page: 001950

 

Transfection Reagent: PEIMAX 32.4 ul/15 cm plate. Packaging Mix: v:g=1:3 (0.1ug/ul). 108 ul/15 cm plate plasmid: pLVX-PEDFR-1 (1.6 ug/ul); 7.2 ug/15 cm plate.

Prepare TR: For one 15 cm plate, in 1ml DMEM(+), add 32.4 ul PEIMAX. Prepare DNA: For one 15 cm plate, in 1 ml DMEM(+), add 108 ul packaging mix and 4.5 ul plasmid (7.2 ug). Add 1ml TR to 1 ml DNA and mix well, keep at room temperature for 20 min, then add it to 15 cm plate drop by drop. 

[[Image:Lentivirus pLVX-PEDFR-1 1day after transfection.JPG]]

1 day after transfection. --[[User:Ztao|Ztao]] 09:25, 18 February 2011 (EST)

[[Category:Lentivirus]]

File:Lentivirus pLVX-PEDFR-1 1day after transfection.JPG

2011-02-18T14:23:06Z

Ztao:

File:Subramanian 1 shRNA 5 comparison of different lentivirus day 1.JPG

2011-02-18T14:19:50Z

Ztao:

File:Subramanian 1 shRNA 4 comparison of different lentivirus day 2.JPG

2011-02-18T14:09:55Z

Ztao:

File:Subramanian 1 shRNA 4 comparison of different lentivirus.JPG

2011-02-17T15:52:36Z

Ztao:

Subramanian 1 shRNA-3

2011-02-16T14:21:48Z

Ztao:

Notebook: Zhen Tao

Page: 001931

 Transfection reagent: PEIMAX, 32.4 ul/flask

Packaging Mix: V:G = 1:3 (prepared by mixing 0.1 ug/ul corresponding DNA). 108 ul/flask

Plasmid (PNPLA2_367881 or PNPLA2_367882): 7.2 ug/flask.

Use 1 ml DMEM(-) to prepare transfection reagent or plasmid, then add transfection reagent to plasmid and keep at room temperature for 20 min; add it to flask drop by drop. 

 

[[Image:Subramanian 1 shRNA 3 PNPLA2-367882-2 PEIMAX day1 after transfection.JPG]]

one day after transfection --[[User:Ztao|Ztao]] 14:41, 11 February 2011 (EST)

[[Image:Subramanian_1_shRNA_3_PNPLA2-367882-2_PEIMAX_day6_after_transfection.JPG]]

6 day after transfection --[[User:Ztao|Ztao]] 09:21, 16 February 2011 (EST)

[[Category:Lentivirus]]

File:Subramanian 1 shRNA 3 PNPLA2-367882-2 PEIMAX day6 after transfection.JPG

2011-02-16T14:20:39Z

Ztao:

File:Subramanian 1 shRNA 3 PNPLA2-367882-2 PEIMAX day1 after transfection.JPG

2011-02-11T19:39:51Z

Ztao:

File:PEDFR to pLVX Plus 2-10-11.JPG

2011-02-10T18:07:58Z

Ztao:

File:Transfection reagent comparison 3 3 day after transfection.JPG

2011-02-10T14:16:16Z

Ztao:

Subramanian 1 shRNA-2

2011-02-10T14:15:41Z

Ztao:

Notebook: Zhen Tao

Page: 001922

 

Transfection reagent: PEIMAX, 32.4 ul/flask

Packaging Mix: V:G = 1:3 (prepared by mixing 0.1 ug/ul corresponding DNA). 108 ul/flask

Plasmid (PNPLA2_216390 or PNPLA2_216192): 7.2 ug/flask.

Use 1 ml DMEM(-) to prepare transfection reagent or plasmid, then add transfection reagent to plasmid and keep at room temperature for 20 min; add it to flask drop by drop.

[[Image:Subramanian 1 shRNA PNPLA2 216390 PEIMAX day1 after transfection.JPG|391x235px|Subramanian 1 shRNA PNPLA2 216390 PEIMAX day1 after transfection.JPG]]

1 day after transfection

--[[User:Ztao|Ztao]] 14:00, 8 February 2011 (EST)

 

[[Image:Subramanian 1 shRNA PNPLA2-216192-1 PEIMAX day2 after transfection.JPG|391x220px|Subramanian 1 shRNA PNPLA2-216192-1 PEIMAX day2 after transfection.JPG]]

[[Image:Subramanian 1 shRNA PNPLA2-216192-2 PEIMAX day2 after transfection.JPG|384x216px|Subramanian 1 shRNA PNPLA2-216192-2 PEIMAX day2 after transfection.JPG]]

[[Image:Subramanian 1 shRNA PNPLA2-216390-1 PEIMAX day2 after transfection.JPG|384x229px|Subramanian 1 shRNA PNPLA2-216390-1 PEIMAX day2 after transfection.JPG]]

[[Image:Subramanian 1 shRNA PNPLA2-216390-2 PEIMAX day2 after transfection.JPG|385x219px|Subramanian 1 shRNA PNPLA2-216390-2 PEIMAX day2 after transfection.JPG]]

2 day after transfection --[[User:Ztao|Ztao]] 11:58, 9 February 2011 (EST)

 

[[Image:Subramanian 1 shRNA PNPLA2-216192-1 PEIMAX day3 after transfection.JPG|381x230px|Subramanian 1 shRNA PNPLA2-216192-1 PEIMAX day3 after transfection.JPG]]

[[Image:Subramanian 1 shRNA PNPLA2-216192-2 PEIMAX day3 after transfection.JPG|379x228px|Subramanian 1 shRNA PNPLA2-216192-2 PEIMAX day3 after transfection.JPG]]

[[Image:Subramanian 1 shRNA PNPLA2-216390-1 PEIMAX day3 after transfection.JPG|379x230px|Subramanian 1 shRNA PNPLA2-216390-1 PEIMAX day3 after transfection.JPG]]

[[Image:Subramanian_1_shRNA_PNPLA2-216390-2_PEIMAX_day3_after_transfection.JPG|378x228px]]

3 day after transfection --[[User:Ztao|Ztao]] 09:15, 10 February 2011 (EST)

[[Category:Lentivirus]]

File:Subramanian 1 shRNA PNPLA2-216390-2 PEIMAX day3 after transfection.JPG

2011-02-10T14:14:33Z

Ztao:

File:Subramanian 1 shRNA PNPLA2-216390-1 PEIMAX day3 after transfection.JPG

2011-02-10T14:13:25Z

Ztao: