Solid Phase DNA is immobilized on a solid support and beads DNA is first

Solid phase dna is immobilized on a solid support and

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Solid Phase – DNA is immobilized on a solid support and beads DNA is first precipitated in the presence of high salt and low pH conditions.
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Precipitated DNA is then adsorbed on to filter membrane (Silica Gel) RNA Extraction Methods – include organic and solid phase methods o MRNA – Can be extracted using immobilized PolyT or PolyU DNA and RNA Concentration o Concentration, yield, and purity assessed using gel analysis, spectrophotometry or fluorometry Buffer – solution of a weak acid and its conjugate base Chapter 6: Proteins Gene – Ordered sequence of nucleotides on a chromosome that encodes a specific functional product Southern Blot – DNA is isolated and cut with restriction enzymes o Fragments are separated by gel electrophoresis, denatured and then transferred to a solid support Electrophoretic Transfer – Uses electric current to move DNA from the gel to membrane Capillary Transfer – Gel is placed on top of a reservoir of buffer, which filter papers soaked in high salt buffer o No instruments required o Procedure is slow Vacuum Transfer – This blotting technique uses suction to move the DNA from the gel to the membrane in a recirculating buffer Northern Blot – modification of Southern blot, designed to investigate RNA Structure and quantity. Western Blot – Immobilized target is protein o Serum, cell lysate or extract is separated on SDS-PAGE DNA Probes – Isolation of a sequence of interest from viral genomes an d in vitro organic synthesis of a piece of nucleic acid RNA Probes – Made by transcription from a synthetic DNA template in vitro Protein Probes – are antibodies that bind specifically to the immobilized target protein Nick Translation – labeled nucleotides are incorporated into single stranded breaks/nicks that are substrates for nucleotide addition by DNA Polymerase Dot Blots – target is deposited in a circle or dot Slot blots – target is deposited in an oblong bar Reverse Dot Blot – Different probes are immobilized on the membrane and the test sample is labeled for hybridization with the immobilized probes. High-Density Oligonucleotide Arrays – used for mutation analysis, single nucleotide polymorphism analysis and sequencing Comparative genome hybridization – designed to test DNA o Used to screen the genome for deletions and amplifications
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