What are the limitations of using an agarose gel to determine if you are homozygous or...

What are the limitations of using an agarose gel to determine if you are homozygous or heterozygous with respect to the D1S80 locus?

Expert Solution

Homozygous : A gene having identical alleles

Heterozygous : A gene having different alleles

D1S80 locus : This locus is found on the petite arm of human chromosome consists of 16 base pairs repeat which are inherited in Mendelian fashion.

Agarose gel electrophoresis : used to determine the sizes of PCR amplified DNA fragments

The main limitations are :

1) we need to prepare 2% agarose gel which solidify very quickly than the other gels like polyacrylamide gel because of high concentration of agarose. So we need to melt when required in a microoven.

2) As the percentage of gel is high it is brittle and may not set uniformly.

3) Agarose gels will not have a uniform pore size.

4) During interpretation we can observe a single band which indicates homozygous at D1S80 locus.

But there is a possibility that two different alleles of nearly equal size can also been intrepreted as homozygous because agarose gel electrophoresis cannot resolve them.

4) If two bands are present it is indicated as heterozygous at D1S80 locus but larger alleles are efficiently amplified than the smaller ones.

5) Need to take care in the preparation of agarose gel this may lead to no bands.

gladiator answered 2 years ago

Write three characteristics that differentiate an agarose gel and a gel polyacrylamide preparation what it is...

Write three characteristics that differentiate an agarose gel and a gel polyacrylamide preparation what it is used for how analyze results

What are the components of agarose gel electrophoresis set up?

When you separate a DNA product of a restriction digest on an agarose gel, and you...

When you separate a DNA product of a restriction digest on an agarose gel, and you see three bands on the gel spread out between the negative and positive poles, what can you say about the DNA, specifically the sizes of the DNA?

3. What is the common label used to detect amplicons if you use agarose gel electrophoresis?...

3. What is the common label used to detect amplicons if you use agarose gel electrophoresis? 5. I ordered an 18-mer oligonucleotide primer from Fisher Scientific. What is an 18-mer? 8. You begin a PCR with 20 copies of the DNA template. After 5 cycles, approximately how many copies of amplicon do you have?

Mary performs a PCR reaction and runs it on an agarose gel, using 100-bp ladder as...

Mary performs a PCR reaction and runs it on an agarose gel, using 100-bp ladder as her molecular weight standard. She images the gel using UV light and sees NO bands (no band in her PCR reaction and no bands for her ladder). Which of the following could explain these results? Explain each answer. 1. Mary forgot to add Gel Red when pouring the gel. 2. Mary's PCR product had an unusually high % of G and C bases. 3....

Linda has asked Brenn to prepare a 1% agarose gel for a gel electrophoresis analysis. But...

Linda has asked Brenn to prepare a 1% agarose gel for a gel electrophoresis analysis. But before he could do that, he needs to prepare 600 ml of 1x TAE from 95x TAE. How much 95x TAE and water should he add to create the 1x TAE solution? Afterwards, how much agarose does he need to weigh out, in grams, and add into 50 ml of 1x TAE?

Lab 9 – Molecular Biology In this lab, you will prepare an agarose gel and use...

Lab 9 – Molecular Biology In this lab, you will prepare an agarose gel and use gel electrophoresis to compare the size of 2 dye molecules Methyl orange and Ponceau G. You will also analyze an “unknown” sample that contains a mixture of two dyes. Dye molecules with lower molecular weight or greater electrical charge will migrate faster through the gel, than dye molecules with greater molecular weights or lesser electrical charge. Materials: Mini gel electrophoresis chamber 6 Tooth comb...

What is the purpose of 1% agarose gel electrophoresis? Why is TAE buffer used during this...

What is the purpose of 1% agarose gel electrophoresis? Why is TAE buffer used during this procedure?

In agarose gel electrophoresis of a sample of DNA, A) small DNA fragments in the sample...

In agarose gel electrophoresis of a sample of DNA, A) small DNA fragments in the sample will move more rapidly through the gel than large DNA fragments. B) the DNA will move through the gel toward the positive electrode. C) the higher the concentration of agarose in the gel, the more rapidly will the DNA move through the gel. D) A and B E) all of the above

Compare and contrast protein (SDS-PAGE) and DNA (agarose) gel electrophoresis.

Question