Ztao at 19:52, 25 March 2011

2011-03-25T19:52:21Z

New page

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]]

3-25-2011 RT-PCR - Revision history

Ztao at 19:52, 25 March 2011