8.8: D1S80 VNTR (Genotyping)
- Page ID
- 24780
\( \newcommand{\vecs}[1]{\overset { \scriptstyle \rightharpoonup} {\mathbf{#1}} } \) \( \newcommand{\vecd}[1]{\overset{-\!-\!\rightharpoonup}{\vphantom{a}\smash {#1}}} \)\(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\) \(\newcommand{\id}{\mathrm{id}}\) \( \newcommand{\Span}{\mathrm{span}}\) \( \newcommand{\kernel}{\mathrm{null}\,}\) \( \newcommand{\range}{\mathrm{range}\,}\) \( \newcommand{\RealPart}{\mathrm{Re}}\) \( \newcommand{\ImaginaryPart}{\mathrm{Im}}\) \( \newcommand{\Argument}{\mathrm{Arg}}\) \( \newcommand{\norm}[1]{\| #1 \|}\) \( \newcommand{\inner}[2]{\langle #1, #2 \rangle}\) \( \newcommand{\Span}{\mathrm{span}}\)\(\newcommand{\AA}{\unicode[.8,0]{x212B}}\)
The minisatellite marker D1S80 is located at 1p35-p36. This VNTR is 16 bases long. With a variation of alleles between 3-24 repeats, the locus displays enough diversity to aid in distinguishing between people. Although this is not a CoDIS marker, the use of multiple loci is required to definitively identify samples. The large repeat (16bp) permits the use of standard agarose gel electrophoresis to explore the diversity of this locus in our lab. PCR products range from 430bp to 814bp long.
- D1S80-for: 5′-GAAACTGGCCTCCAAACACTGCCCGCCG-3′
- D1S80-rev: 5′-GTCTTGTTGGAGATGCACGTGCCCCTTGC-3′
- PCR the DNA samples extracted from cheek cells using the PCR Beads.
- Pour 2% agarose into casting apparatus in the refrigerator.
- 2 gels per class need to be made → 100ml of TBE with 2g agarose
- Add 5μl SYBR safe solution into the molten agarose before casting.
- Place 2 sets of combs into the gel → at one end and in the middle
- Load the gel with a DNA ladder.
- The sample is from the PCR.
- Run the gel at 120V for 20 minutes.
- Visualize on the UV transilluminator.
Example Results
1. How many alleles are visible in each lane?
2. Are the genotypes distinguishable between individuals?
3. Are any of the alleles common between individual samples?