Why does a smaller DNA fragment move faster than a larger one?
DNA fragments are negatively charged, so they move towards the positive electrode. Because all DNA fragments have the same amount of charge per mass, small fragments move through the gel faster than large ones.
Why is it necessary to have a primer on each side of the DNA sequence to be amplified?
1. Why is it necessary to have a primer on each side of the DNA segment to be amplified? The primer is able to mark the spot where Taq polymerase must make matching strands.
What is needed from the cells for PCR?
and the PCR will not occur. What is needed from the cells for PCR? The genomic DNA is required for the polymerase chain reaction to occur since PCR amplifies the DNA. sequences.
Why is the DNA stored cold with the Instagene matrix after boiling the samples?
The DNA is stored cold with the instagene matrix after boiling samples in order to slow bacterial growth. In that way, you will greatly slow down the activity of any remaining enzymes that could harm your DNA. Most likely it is a way of preserving the DNA and to avoid it bacterial contamination.
What is the purpose of InstaGene Matrix added to DNA?
InstaGene matrix, made with a specially formulated 6% w/v Chelex resin, makes DNA sample preparation fast, easy, and cost-effective, providing PCR-quality template DNA in less than an hour. The Chelex matrix binds to PCR inhibitors rather than DNA, preventing DNA loss due to irreversible DNA binding.
What are the three steps in a PCR cycle and what does each step accomplish?
PCR is based on three simple steps required for any DNA synthesis reaction: (1) denaturation of the template into single strands; (2) annealing of primers to each original strand for new strand synthesis; and (3) extension of the new DNA strands from the primers.
How many primers are used in RT-PCR?
Which enzyme is used in RT-PCR?
enzyme reverse transcriptase
How much cDNA do you need for real time PCR?
In practice, this is dependent on your gene of interest ofcourse, and many other factors that Elena eluted to, but just aim for using ~g of total RNA for cDNA synthesis, with your final cDNA volume ~20-50ul, and use 1ul for PCR.