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alycja 02-07-2013 08:35 PM

comparative qPCR - no melting curve
:help: 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?

alycja 06-05-2013 04:43 PM

Re: comparative qPCR - no melting curve

Originally Posted by qianhaha (Post 447359)
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.

:wacko::unsure: How is this info relevant?????

luisillo 06-06-2013 01:25 PM

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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