the common baker's yeast, Saccharomyces cerevisiae, contains about 7000 genes; with a microarray, one can measure qualitatively how each gene is expressed, and how that expression changes, for example, with a change in temperature. There are many different ways to fabricate microarrays; the most common are silicon chips, microscope slides with spots of ~ 100 micrometre diameter. There can be anywhere from 100 spots to more than 10,000 on a given array. Arrays can also be made with molecules other than DNA. For example, an antibody array can be used to determine what proteins or bacteria are present in a blood sample. Allele Specific Oligonucleotide Allele specific oligonucleotide (ASO) is a technique that allows detection of single base mutations without the need for PCR or gel electrophoresis. Short (20-25 nucleotides in length), labeled probes are exposed to the non- fragmented target DNA. Hybridization occurs with high specificity due to the short length of the probes and even a single base change will hinder hybridization. The target DNA is then washed and the labeled probes that didn't hybridize are removed. The target DNA is then analyzed for the presence of the probe via radioactivity or fluorescence. ragi roti umum itu, Saccharomyces cerevisiae, mengandung sekitar 7000 gen, dengan microarray dapat mengukur secara kualitatif bagaimana setiap gen diekspresikan, dan Page 8
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the common baker's yeast, Saccharomyces cerevisiae, contains
about 7000 genes; with a microarray, one can measure qualitatively
how each gene is expressed, and how that expression changes, for
example, with a change in temperature. There are many different
ways to fabricate microarrays; the most common are silicon chips,
microscope slides with spots of ~ 100 micrometre diameter. There
can be anywhere from 100 spots to more than 10,000 on a given
array. Arrays can also be made with molecules other than DNA. For
example, an antibody array can be used to determine what proteins
or bacteria are present in a blood sample.
Allele Specific Oligonucleotide
Allele specific oligonucleotide (ASO) is a technique that allows
detection of single base mutations without the need for PCR or gel
electrophoresis. Short (20-25 nucleotides in length), labeled probes are
exposed to the non-fragmented target DNA. Hybridization occurs with
high specificity due to the short length of the probes and even a single
base change will hinder hybridization. The target DNA is then washed
and the labeled probes that didn't hybridize are removed. The target
DNA is then analyzed for the presence of the probe via radioactivity
or fluorescence.
ragi roti umum itu, Saccharomyces cerevisiae, mengandung sekitar
7000 gen, dengan microarray dapat mengukur secara kualitatif
bagaimana setiap gen diekspresikan, dan bagaimana perubahan
ekspresi gen, misalnya, akibat perubahan suhu. Ada banyak cara
untuk membuat microarray, yang paling umum adalah chip silikon,
mikroskop slide dengan diameter bintik dari 100 mikrometer. Ada
juga diameter bintik yang lebih dari 10.000 mikrometer. Arrays juga
bisa dibuat dengan molekul lain dari DNA. Sebagai contoh, sebuah
array antibodi dapat digunakan untuk menentukan kehadiran protein
atau bakteri dalam sampel darah.
Alel spesifik oligonukleotida
Alel spesifik Oligonukleotida (ASO) adalah teknik yang
memungkinkan deteksi mutasi basa tunggal tanpa memerlukan
elektroforesis PCR atau gel. Gelombang pendek (20-25 nukleotida),
probe berlabel terkena DNA target tidak terfragmentasi. Hibridisasi
terjadi dengan spesifisitas yang tinggi terhadap panjang gelombang
yang pendek dari probe meskipun terjadi perubahan basa tunggal
yang akan menghambat hibridisasi. DNA target kemudian dicuci
dan probe label yang tidak berhibridisasi dihapus. DNA target
kemudian dianalisa untuk mengetahui probe melalui radioaktivitas