Helpful tips

What is stem loop primer?

What is stem loop primer?

The RT-primer contains a highly stable stem-loop structure that lengthens the target cDNA. The forward PCR primer adds additional length with nucleotides that optimize its melting temperature (T(m)) and enhance assay specificity. The reverse primer disrupts the stem loop.

What is stem loop PCR?

Stem-loop quantitative reverse transcription PCR (RT-qPCR) is a molecular technique used for identification and quantification of individual small RNAs in cells. Identification of the two small RNAs was carried out by sequencing the PCR-amplified small RNA products upon ligation into the pJET1. 2/blunt vector.

How do you design a primer for miRNA?

  1. Open miRBase, enter interested sequence.
  2. Open respective miRNA & fetch the Sequence.
  3. Alongside, open the miRNA Primer Design Tool & login in that software.
  4. Enter the miRNA sequence in the box & select URL probe.
  5. Enter the design option.
  6. select the Universal reverse & forward primer according to your convenience.

What is reverse transcriptase PCR used for?

The Polymerase Chain Reaction Reverse transcription (RT)-PCR is used to amplify RNA targets. The RNA template is converted into complementary (c)DNA by the enzyme reverse transcriptase. The cDNA serves later as a template for exponential amplification using PCR. RT-PCR can be undertaken in one or two steps.

What is universal reverse primer?

The universal reverse primer is designed for use with the Human U6 forward primer provided in our High-Specificity miRNA QRT-PCR Detection Kit for a complete, validated assay requiring no upfront design work. The Kit delivers sensitive and reproducible detection of miRNA with high specificity.

What is the seed region of miRNA?

What is the “seed sequence” of a miRNA? miRNAs regulate the gene expression by binding to the mRNA. The seed sequence is essential for the binding of the miRNA to the mRNA. The seed sequence or seed region is a conserved heptametrical sequence which is mostly situated at positions 2-7 from the miRNA 5´-end.

How many primers are used in each PCR reaction?

Two primers
Two primers are used in each PCR reaction, and they are designed so that they flank the target region (region that should be copied).