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annealing-temperature

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Journal Articles
The American Biology Teacher. 2012; 744256–260 doi: https://doi.org/10.1525/abt.2012.74.4.9
Published: 01 April 2012
.... In this molecular biology laboratory, students learn the steps of PCR with an emphasis on primer composition and annealing temperature, which they manipulate to test the effect on successful DNA amplification. Students design experiments to test their hypotheses, promoting a discovery-based approach...
Journal Articles
The American Biology Teacher. 2020; 82149–51 doi: https://doi.org/10.1525/abt.2020.82.1.49
Published: 01 January 2020
... deoxyribonucleotides (dNTPs), primers (short oligonucleotides that define the amplified sequence), and thermostable DNA polymerase ( Garibyan & Avashia, 2013 ). During the reaction, three temperature steps are repeated (cycling; usually 25–30 times): denaturation (95°C; DNA strands are separated), annealing (55–65...
Images
Published: 01 April 2012
Figure 2. Typical patterns of PCR product analysis by ethidium bromide staining of agarose gels produce one band of amplified cDNA for each set of (A) claudin-2 primers and (B) claudin-12 primers when using annealing temperature ranging from 51°C to 72°C. Figure 2. Typical patterns of PCR
Journal Articles
The American Biology Teacher. 2017; 799763–768 doi: https://doi.org/10.1525/abt.2017.79.9.763
Published: 01 November 2017
... effect of varying annealing temperatures on the resulting PCR product (i.e., low temperatures may lead to nonspecific amplification, and high temperatures may lead to lack of amplification). To determine if the COI region was successfully amplified, students used agarose gel electrophoresis to detect...
Journal Articles
The American Biology Teacher. 2017; 799702–710 doi: https://doi.org/10.1525/abt.2017.79.9.702
Published: 01 November 2017
... name suggests cycles through various temperatures. The biological samples collected from the Worthington crime scene have been taken to the lab for further analysis. The first step of making a DNA profile is to extract DNA from all of the crime scene samples. Today you will practice extracting DNA...
Images
Published: 01 April 2012
have the rotor balanced. They were also asked to look at the settings from a PCR machine and state the annealing temperature and time required. The success rate of the students on these questions was analyzed. Bars represent the mean ± SE (*P < 0.05). Data are from both classes analyzed separately
Journal Articles
The American Biology Teacher. 2015; 772118–125 doi: https://doi.org/10.1525/abt.2015.77.2.7
Published: 01 February 2015
... precautions should be observed: a Wear gloves. b Wear eye protection when pouring hot solutions. c High voltage; disconnect power before opening chambers. d Thermal cycler components reach high temperatures during PCR cycling. Suggested vendors are listed in a file of Supplemental Materials...
Journal Articles
The American Biology Teacher. 2014; 765338–344 doi: https://doi.org/10.1525/abt.2014.76.5.8
Published: 01 May 2014
... denatured to separate the DNA into two separate strands. Next the temperature is lowered so that the primers can anneal (or attach) to the DNA strands only in the microsatellite region of interest. Finally, the temperature is raised so that the enzyme DNA polymerase can elongate (synthesize or build) a new...
Journal Articles
The American Biology Teacher. 2013; 759704–707 doi: https://doi.org/10.1525/abt.2013.75.9.13
Published: 01 November 2013
... with their own tissue absorbed students in this 2-hour practical. Table 1. PCR was run as follows. Steps 2a to 2c were repeated 35 times. Step Temperature (°C) Time 1. Initial denaturation 95 5 minutes 2a. Denaturation 95 20 seconds 2b. Annealing 60 20 seconds 2c...
Journal Articles
The American Biology Teacher. 2012; 749667–671 doi: https://doi.org/10.1525/abt.2012.74.9.14
Published: 01 November 2012
... BLAST.......................5:318-I Boiling point.......................5:343-Q Annealing temperature.......................4:256-H Antique microscope slide.......................5:311-A, 5 Applied ecology.......................7:445-A Bacteria.......................5:305-F...
Journal Articles
The American Biology Teacher. 2012; 744232–236 doi: https://doi.org/10.1525/abt.2012.74.4.5
Published: 01 April 2012
... have the rotor balanced. They were also asked to look at the settings from a PCR machine and state the annealing temperature and time required. The success rate of the students on these questions was analyzed. Bars represent the mean ± SE (*P < 0.05). Data are from both classes analyzed separately...
Journal Articles
The American Biology Teacher. 2011; 736331–335 doi: https://doi.org/10.1525/abt.2011.73.6.5
Published: 01 August 2011
... not available, one could be borrowed from a neighboring university or community college. A commonly used cycling program is 30–35 cycles of the following temperatures for the indicated periods: 95°C for 1 minute (DNA melting/denaturation step), 55°C for 1 minute (primer annealing step), and 72°C for...
Journal Articles
The American Biology Teacher. 2008; 707411–414 doi: https://doi.org/10.2307/30163313
Published: 01 September 2008
... to avoid the cell debris and Chelex beads at the bottom of the tube. 10. Store the samples in the freezer or on ice if continuing with the DNA amplification. 6. Amplify the DNA samples using the following PCR cycle for 30 cycles: . Denaturing time & temperature: 30 seconds at 94 o C . Annealing time...
Journal Articles
The American Biology Teacher. 2007; 695280–286 doi: https://doi.org/10.2307/4452157
Published: 01 May 2007
... several temperature steps that trigger the denaturing of the double helix, the annealing of primers to target DNA sequences, and an extension of new strands with deoxy- nucleotides. The result is a new identical copy of the desired DNA fragment as shown in Figure 3. The process is repeated until DNA...
Journal Articles
The American Biology Teacher. 2004; 663183–189 doi: https://doi.org/10.2307/4451652
Published: 01 March 2004
... the class should have worked but may not have been optimum because of a disparity in the ideal annealing temperatures of the two primers (50?C and 58C). The project was a success in that the students gained experience with DNA extraction, PCR, and gel electrophoresis. They also learned a great deal...
Journal Articles
The American Biology Teacher. 2000; 62151–57 doi: https://doi.org/10.2307/4450826
Published: 01 January 2000
...- ever, since the DNA fragment that we excised from the gel has the entire RAPD primer at each end, we can run the reaction at high stringency. We used a 450 C annealing temperature, no slow temperature ramp, and only 25 cycles to produce the single band shown in Figure 3, Arrow A. We then used six...
Journal Articles
The American Biology Teacher. 2001; 638596–605 doi: https://doi.org/10.2307/4451191
Published: 01 October 2001
.../HindIII Markers (G1711) - aliquot 10 gl for each group. Have students heat at 65 C for 5 min- utes to disrupt cohesive ends that may anneal before loading gel. *At the time of press these vectors were dis- continued by Promega. Identical vectors are available by contacting Promega, or pET-5a and pET-9a...
Journal Articles
The American Biology Teacher. 1998; 604276–280 doi: https://doi.org/10.2307/4450471
Published: 01 April 1998
... match at the 3'-terminus is required for good amplification. It is advisable to select the length and loca- tion of the primer on the target DNA so that the annealing temperature for each primer is high (60-70? C). Generally, primers should have a G + C content of about 50% with a random base dis...
Journal Articles
The American Biology Teacher. 1994; 568476–481 doi: https://doi.org/10.2307/4449892
Published: 01 November 1994
... the number of copies of DNA occurs, requiring only about two and one-half hours for production of many millions of copies. Reaction temperatures for PCR do not vary much for amplification of nucleic acids from various organisms, except for annealing temperature. As Table 1 shows, denaturing and ex...
Journal Articles
The American Biology Teacher. 1998; 606448–452 doi: https://doi.org/10.2307/4450519
Published: 01 June 1998
... to one hour at room temperature). It is very important to make sure the gel solidifies completely before removing the comb. If the comb is removed too early, the wells may deform, and the RFLP LAB EXERCISE 449 | 1 2 3l 23,130 32,710 23,130- 21,230 9416- -10,630 6557 ~ ~7420 6557-- -5800, 5650 4361...