UV or blue light is used to excite the fluorescence of the DNA in the gel. Typically fluorescence arises from intercalating dyes such as SYBR Green that bind to DNA during electrophoresis.
Also called thermocyclers, these machines control the temperature environment of the PCR reaction. In a traditional PCR reaction, 3 temperatures are used to cycle the sample through steps of denaturation, annealing, and extension.
These are thin-walled, low-volume tubes designed for efficient heat transfer. The SLIM Cycler, Mini8, and T-8 all use standard 200 microliter PCR tubes.
dNTPs (deoxynucleotides) are called bases and are the building blocks of DNA: the A, T, C and G. A and T are complementary, as are C and G. When the complementary bases come together, they are called a base pair.
PCR works best under very specific environmental conditions, including specific chemistries and pH. For example, PCR buffers usually include magnesium ions and are kept at 8.4 pH.
Synthesized single stranded DNA usually around 20 base pairs long. The sequence of these primers are the exact complement to the segment of the template DNA sequence to be copied.
A DNA sequence to be targeted and copied by PCR. The number of base pairs (or length) of the amplified product can vary from ~50 bases to thousands. By the way, the human genome is 3 billion base pairs long!
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Slim Cycler Student Lab - Who killed Carol?
In this 90-minute lab, you will help solve Carol’s murder case by assisting the forensic investigators in analyzing numerous DNA samples from the scene of the crime using molecular biology techniques such as polymerase chain reaction (PCR) and gel electrophoresis.
This exercise will familiarize students with molecular biology techniques used in real-world applications such as DNA profiling in a crime scene investigation.
Aas develops PCR curriculums with the goal of providing students with genetic tools they can use to solve real world problems that involves DNA profiling. The information below will allow students to conduct state-of-the-art molecular biology experiments and understand how these techniques are performed in the real world forensic science labs.
Following PCR amplification of DNA, agarose gel electrophoresis is commonly used to separate DNA by size. By running a sample with a DNA ladder of known DNA lenghts, you can determine the length of the amplified product.
Micropipettes and tips
These common lab tools are used to manipulate microliters (1/1000 of a milliliter) of fluid and is very useful in almost any wet lab.
Other PCR lab supplies
- Scale (to weigh agarose)
- Latex gloves (to protect yourself and your sample)
- Gel tray and combs
- Microtest tubes and rack
- Electrophoresis buffer
- DNA staining agent
- Gel loading buffer
- DNA ladder or marker
The enzyme that brings in free floating nucleotides to attach to the primer and extend the copied sequence. Imagine if a zipper's chains are the DNA, then polymerase would be the slider. Taq polymerase is commonly used.
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