|
|
|
|
| LEADER |
01000caa a22002652 4500 |
| 001 |
NLM120958406 |
| 003 |
DE-627 |
| 005 |
20250203115619.0 |
| 007 |
tu |
| 008 |
231222s2002 xx ||||| 00| ||eng c |
| 028 |
5 |
2 |
|a pubmed25n0403.xml
|
| 035 |
|
|
|a (DE-627)NLM120958406
|
| 035 |
|
|
|a (NLM)12238766
|
| 040 |
|
|
|a DE-627
|b ger
|c DE-627
|e rakwb
|
| 041 |
|
|
|a eng
|
| 100 |
1 |
|
|a Chiou, Jeffrey T
|e verfasserin
|4 aut
|
| 245 |
1 |
0 |
|a Thirty-cycle temperature optimization of a closed-cycle capillary PCR machine
|
| 264 |
|
1 |
|c 2002
|
| 336 |
|
|
|a Text
|b txt
|2 rdacontent
|
| 337 |
|
|
|a ohne Hilfsmittel zu benutzen
|b n
|2 rdamedia
|
| 338 |
|
|
|a Band
|b nc
|2 rdacarrier
|
| 500 |
|
|
|a Date Completed 07.05.2003
|
| 500 |
|
|
|a Date Revised 10.12.2019
|
| 500 |
|
|
|a published: Print
|
| 500 |
|
|
|a Citation Status MEDLINE
|
| 520 |
|
|
|a The performance of a novel thermal cycler has been characterized in a 30-cycle PCR. The device consists of a microcapillary equipped with bidirectional pressure-driven flow and in situ optical position sensors. A 1-microL droplet of reaction mixture moves between three heat zones in a 1-mm i.d., oil-filled capillary using a multi-element scattered light detector and active feedback. The design permits time and number of cycles to be changed without hardware modification, unlike other flow-in-capillary PCR systems. Temperature optimization has been performed on the three PCR heat steps. The optimal denaturation temperature is 94 degrees C-96 degrees C, which is identical to commercial machines. The optimal extension temperature of 62 degrees C-66 degrees C is lower than reported for Taq DNA polymerase (70 degrees C-80 degrees C) because of the high enzyme concentration and/or the absence of detergent in the PCR mixture. The optimal annealing temperature seems to be the same as the optimal extension temperature. This is because extension occurs when the sample is inside of the annealing heat zone. Annealing takes place as the sample travels between heat zones. Device speed (23 minfor 30 cycles without time optimization) is competitive with other rapid PCR designs for efficiencies comparable to a commercial machine
|
| 650 |
|
4 |
|a Comparative Study
|
| 650 |
|
4 |
|a Evaluation Study
|
| 650 |
|
4 |
|a Research Support, U.S. Gov't, Non-P.H.S.
|
| 650 |
|
4 |
|a Research Support, U.S. Gov't, P.H.S.
|
| 650 |
|
4 |
|a Technical Report
|
| 650 |
|
4 |
|a Journal Article
|
| 650 |
|
7 |
|a DNA Primers
|2 NLM
|
| 650 |
|
7 |
|a DNA, Bacterial
|2 NLM
|
| 650 |
|
7 |
|a Taq Polymerase
|2 NLM
|
| 650 |
|
7 |
|a EC 2.7.7.-
|2 NLM
|
| 700 |
1 |
|
|a Matsudaira, Paul T
|e verfasserin
|4 aut
|
| 700 |
1 |
|
|a Ehrlich, Daniel J
|e verfasserin
|4 aut
|
| 773 |
0 |
8 |
|i Enthalten in
|t BioTechniques
|d 1993
|g 33(2002), 3 vom: 11. Sept., Seite 557-8, 560, 562 passim
|w (DE-627)NLM012627046
|x 0736-6205
|7 nnns
|
| 773 |
1 |
8 |
|g volume:33
|g year:2002
|g number:3
|g day:11
|g month:09
|g pages:557-8, 560, 562 passim
|
| 912 |
|
|
|a GBV_USEFLAG_A
|
| 912 |
|
|
|a SYSFLAG_A
|
| 912 |
|
|
|a GBV_NLM
|
| 912 |
|
|
|a GBV_ILN_21
|
| 912 |
|
|
|a GBV_ILN_22
|
| 912 |
|
|
|a GBV_ILN_39
|
| 912 |
|
|
|a GBV_ILN_40
|
| 912 |
|
|
|a GBV_ILN_50
|
| 912 |
|
|
|a GBV_ILN_60
|
| 912 |
|
|
|a GBV_ILN_62
|
| 912 |
|
|
|a GBV_ILN_65
|
| 912 |
|
|
|a GBV_ILN_70
|
| 912 |
|
|
|a GBV_ILN_99
|
| 912 |
|
|
|a GBV_ILN_121
|
| 912 |
|
|
|a GBV_ILN_130
|
| 912 |
|
|
|a GBV_ILN_227
|
| 912 |
|
|
|a GBV_ILN_350
|
| 912 |
|
|
|a GBV_ILN_618
|
| 912 |
|
|
|a GBV_ILN_640
|
| 912 |
|
|
|a GBV_ILN_754
|
| 912 |
|
|
|a GBV_ILN_2001
|
| 912 |
|
|
|a GBV_ILN_2002
|
| 912 |
|
|
|a GBV_ILN_2003
|
| 912 |
|
|
|a GBV_ILN_2005
|
| 912 |
|
|
|a GBV_ILN_2006
|
| 912 |
|
|
|a GBV_ILN_2007
|
| 912 |
|
|
|a GBV_ILN_2008
|
| 912 |
|
|
|a GBV_ILN_2009
|
| 912 |
|
|
|a GBV_ILN_2010
|
| 912 |
|
|
|a GBV_ILN_2012
|
| 912 |
|
|
|a GBV_ILN_2015
|
| 912 |
|
|
|a GBV_ILN_2018
|
| 912 |
|
|
|a GBV_ILN_2023
|
| 912 |
|
|
|a GBV_ILN_2035
|
| 912 |
|
|
|a GBV_ILN_2040
|
| 912 |
|
|
|a GBV_ILN_2060
|
| 912 |
|
|
|a GBV_ILN_2099
|
| 912 |
|
|
|a GBV_ILN_2105
|
| 912 |
|
|
|a GBV_ILN_2121
|
| 912 |
|
|
|a GBV_ILN_2470
|
| 951 |
|
|
|a AR
|
| 952 |
|
|
|d 33
|j 2002
|e 3
|b 11
|c 09
|h 557-8, 560, 562 passim
|