6.
6.1.
- 300 nM ;
( TaqMan) - 250 nM;
MgCl2 - 5 nM (SYBR Green), 3.5 nM (TaqMan);
- AmpliTaqGold, Applied Biosystems.
6.2. SYBR Green
AmpliTaqGold, Applied Biosystems
| | |
1 | 8 min | 94OC |
2 | 30 sec | 94OC |
3 | 20 sec | 60-65OC |
4 | 1 min | 72OC |
5 | Plate read |
6 | Goto 2 for 39 times |
7 | Melting curve from 60OC to 94OC read every 0.2OC hold for 1 sec before plate read |
8 | Forever | 10OC |
|
- , , . , 15-20 , .
- (Plate read) 72OC, ( . ).
- SYBR Green, , C(T) 35-38, 40 .
- AmpliTaqGold 60OC, . 3 4 1 min & 60-65OC, .
- (melting curve) SYBR Green PCR. , PCR- .
6.3. TaqMan
AmpliTaqGold, Applied Biosystems
| | |
1 | 10 min | 94OC |
2 | 15 sec | 94OC |
4 | 1 min | 60OC |
5 | Plate read |
6 | Goto 2 for 39-45 times |
8 | Forever | 10OC |
|
- Applied Biosystems -N-, dTTP dUTP, PCR-. -N-, 2 min 50OC.
- (, PCR-, ). , . , , .
|
6. |
|
, ,
Re /21.06.2006 15:44/
Mg? nM mM?
Cinderella /14.06.2007 08:58/
? ?
Guest /14.06.2007 10:51/
- ?
/14.06.2007 11:10/
(Cinderella @ 14.06.2007 08:58) ? ? +(383) 3326636, 334710: +(383) 3326636e-mail medigen@sibmail.ru : +(495) 113 20 52 /05.02.2008 16:38/
!!! ? 200, 1 . . . ( ) , , 3000 .
volmin /17.11.2019 04:30/
watchesbiz /03.11.2021 17:18/
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