6. Annealing Temperature (Ta): • The primer melting temperature is the estimate of the DNA-DNA hybrid stability and critical in determining the annealing temperature. Depends directly on length and GC composition of the primers. . • Too high Ta - produce insufficient primer-template hybridization. • Too low Ta - lead to non-specific products caused by a high number of base pair mismatches. 47/49 Now you should be able to answer the following questions >> How the amplification will be done? » How you will determine your target sequence? >> How the amplification will be specific for certain segment? >> What are the requirements to carry PCR?

Human Anatomy & Physiology (11th Edition)
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ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
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Chapter1: The Human Body: An Orientation
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Draw one cycle of PCR reaction below the following diagram
16:46 865
.
6. Annealing Temperature (Ta):
• The primer melting temperature is the estimate of the DNA-DNA hybrid stability and critical in
determining the annealing temperature.
• Depends directly on length and GC composition of the primers.
>>
Mol_Biol_Tech_le...
• Too high Ta - produce insufficient primer-template hybridization.
Too low Ta - lead to non-specific products caused by a high number of base pair mismatches.
>>
>>>
Primer Design Guidelines
>>
Now you should be able to answer the following questions
How the amplification will be done?
How you will determine your target sequence?
How the amplification will be specific for certain
segment?
What are the requirements to carry PCR?
|||
A. Draw one cycle of PCR reaction below the following diagram.
B. Label the template DNA, the primers, and what is happening at each step.
(19
XXXX
Homework
• Suppose you perform a PCR that begins with one double-strand of the
following DNA template:
→5'-CTACCTGCGGGTTGACTGCTACCTTCCCGGGATGCCCAAAATTCTCGAG-3-
+3'-GATGGACGCCCAACTGACGATGGAAGGGCCCTACGGGTTTTAAGAGCTC-5'+
(O)
1
@alball ■
Save:
température
2
cycle #1
(3)
47/49
<
48
48/49
49/49
Transcribed Image Text:16:46 865 . 6. Annealing Temperature (Ta): • The primer melting temperature is the estimate of the DNA-DNA hybrid stability and critical in determining the annealing temperature. • Depends directly on length and GC composition of the primers. >> Mol_Biol_Tech_le... • Too high Ta - produce insufficient primer-template hybridization. Too low Ta - lead to non-specific products caused by a high number of base pair mismatches. >> >>> Primer Design Guidelines >> Now you should be able to answer the following questions How the amplification will be done? How you will determine your target sequence? How the amplification will be specific for certain segment? What are the requirements to carry PCR? ||| A. Draw one cycle of PCR reaction below the following diagram. B. Label the template DNA, the primers, and what is happening at each step. (19 XXXX Homework • Suppose you perform a PCR that begins with one double-strand of the following DNA template: →5'-CTACCTGCGGGTTGACTGCTACCTTCCCGGGATGCCCAAAATTCTCGAG-3- +3'-GATGGACGCCCAACTGACGATGGAAGGGCCCTACGGGTTTTAAGAGCTC-5'+ (O) 1 @alball ■ Save: température 2 cycle #1 (3) 47/49 < 48 48/49 49/49
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