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Normalization for RNA or cDNA during two step RT …

In this more streamlined version of the Two-Cycle protocol, the first-cycle cDNA cleanup step is removed since the amount of material is very limiting and it has been shown that omitting this step does not affect the final array results.

ReadyScript cDNA Synthesis Mix is a sensitive and easy-to-use solution for two-step RT-PCR

Hello :-)

Excuse my stupid questions.

About this RNA normalization..... Here I was given an RNA quality control protocol, you can see it.

1. What is this RNA normalization?

2. How someone could normalize the amount of mRNA if it not visible on the gel, as seen in this protocol?

3. Apart from how intact is my RNA, what more information gives me the RNA electrophoresis, if I see only the rRNA?

4. If I load my RNA sample on a 1% agarose and I don`t see anything on the level of the upper marker, would it mean that the sample is not being contaminated with gDNA?

5. As far as I undestand, non-PCR-ed gDNA makes something like.....bands with approximate size at about.....8000-10 000 bp, is that correct?

Any answer will be highly appreciated,

Nephrite

dART RT-PCR Kiti / cDNA Sentez Kiti (Two-Step) - …

In Chapter 10, we describe two methods of complementary DNA (cDNA) synthesis and amplification

Although one-step RT-PCR offers simplicity and convenience and minimizes the possibility for contamination, the resulting cDNA cannot be repeated used as in two step RT-PCR.

In the Two-Cycle Target Labeling protocol, the first-cycle cDNA cleanup step is omitted. Is there any impact on the data by removing this step?

Two-Step RT-PCR Kit; Product Name Size Cat

Perform cDNA synthesis in two step real time quantitative RT-PCR (RT-qPCR) applications with the convenient, optimized system of First Strand cDNA Synthesis …

First-strand cDNA Synthesis Kit—qSTAR first-strand cDNA synthesis kit for miRNA is a two-step protocol kit.
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