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comparative qPCR - no melting curve

comparative qPCR - no melting curve - Biology Forum

comparative qPCR - no melting curve - Biology Forums. Ask questions and discuss the study of Biology. If you have Biology questions from your homework ask them here!


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Old 02-07-2013, 08:35 PM
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Question comparative qPCR - no melting curve



Hi!
I'm performing qPCR with Roche SYBR green Fast Start (Before I used Bio-Rad and everything worked).
I added disocciation curve in tem 52C, my primers melt at 54C, my reference gene in 57C.
My reference gene showed nice disocciation curve peak, BUT NOT any of my genes! No curve at alll of them (cDNA, no-RT, NTC). The Ct value of amplification looks very nice for cDNA, for NTC there is no amplification... just this lack of melting curves seems very strange. What could possibly went wrong?
Thanks
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  #2  
Old 06-05-2013, 04:43 PM
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Question Re: comparative qPCR - no melting curve

Quote:
Originally Posted by qianhaha View Post
In organic chemistry, peptide synthesis is the production of peptides, which are organic compounds in which multiple amino acids are linked via amide bonds which are also known as peptide bonds. The biological process of producing long peptides (proteins) is known as protein biosynthesis. We use peptide 2.0 Inc</a> for peptide synthesis. Synthesized peptides are used in applications such as designing enzymes, testing drugs, and creating antibodies. Rather than synthesizing their own peptides, many scientists outsource the job to custom services. There are two main avenues of peptide synthesis: solid-phase or liquid-phase synthesis. In the more common solid-phase synthesis, the C-terminus is protected by attachment to a solid resin, which also simplifies separating the peptide from the reaction mixture. Liquid-phase synthesis, or synthesis in solution, is slower and labor-intensive, but has the advantages of multiple rounds of purification, and the opportunity for convergent synthesis, in which synthesized peptides can be attached to form larger ones. Options available from most custom synthesis services include: design of the peptide with labels or modifications (such as phosphorylation, methylation, biotinylation, glycosylation, cyclization, or attachment to carrier proteins or dye labels); quantity; purification (most are purified by HPLC); verifiable purity (most are analyzed by mass spectrometry and/or analytical HPLC); and solubility testing.
How is this info relevant?????
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  #3  
Old 06-06-2013, 01:25 PM
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Default Re: comparative qPCR - no melting curve

It is not. This qianhaha must be a spammer since he/she has been posting the same thing over and over. I think is above 20 times for what I have seen.
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