random Protocols

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

Shotgun sequencing of a large segment of DNA involves random fragmentation of the target region into smaller segments that are subsequently cloned into a bacteriophage M13 vector. The goal is to create a library of overlapping clones that provide at least fivefold coverage over the entire length of the target fragment. - [Read Generation of a Library of Randomly Overlapping DNA Inserts Protocol]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to obtain progeny as random ascospore isolates. - [Read How to Obtain Progeny as Random Ascospore Isolates]

Category: Cloning Protocols: Mutagenesis Protocols

Protocol for hydroxylamine mutagenesis of plasmid DNA is ideal for random mutagenesis of plasmid DNA which is then used in a plasmid shuffle or screen for ts mutants. - [Read Hydroxylamine Mutagenesis of Plasmid DNA Protocol]

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

Protocol for in vitro transcription and translation using the coupled reticulocyte lysate system. This protocol is designed to test random samples on a protein gel. Scale up the reactions accordingly. Protocol includes: Procedure, Solutions, BioReagents and Chemicals and protocol hints. - [Read In Vitro Transcription and Translation Using the Coupled Reticulocyte Lysate System]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Procedures for labeling DNA, RNA, and oligonucleotides with DIG, biotin, or fluorochromes. Includes: Random primed labeling of ds DNA with DIG-, Biotin- or Fluorescein-High Prime reaction mix; PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes. - [Read Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes]

Category: Nucleic Acid Modification Protocols: DIG Biotin Labeled Probes Protocols

Procedures for labeling DNA, RNA, and oligonucleotides with DIG, Biotin, or Fluorochromes. Includes: Random primed labeling of ds DNA with DIG-, Biotinor Fluorescein-High Prime reaction mix; PCR labeling of ds DNA with the PCR DIG Probe Synthesis Kit or PCR Labeling Mixes. - [Read Procedures for Labeling DNA, RNA, and Oligonucleotides with DIG, Biotin, or Fluorochromes]

Category: Nucleic Acid Modification Protocols: Nucleic Acid Radiolabeling Protocols

Protocol for radiolabeling of DNA by extension of random oligonucleotides in the presence of melted agarose. - [Read Radiolabeling of DNA by Extension of Random Oligonucleotides in the Presence of Melted Agarose]

Category: Nucleic Acid Modification Protocols: Nucleic Acid Radiolabeling Protocols

In this procedure, synthesis of cDNA is carried out in the presence of saturating concentrations of all four dNTPs and trace amounts of a single radiolabeled dNTP.  After subtraction hybridization, the enriched single-stranded cDNA is radiolabeled to high specific activity in a second synthetic reaction by extension of random oligonucleotide primers using the Klenow fragment of E. coli DNA polymerase. - [Read Radiolabeling of Subtracted cDNA Probes by Random Oligonucleotide Extension Protocol]

Category: Fungi Protocols: Aspergillus Protocols

RAPD is a procedure for typing and fingerprinting isolates of a species. It can be used for epidemiological studies, such as investigations into hospital outbreaks and as a laboratory aid to keep track of cultures and to verify that mutants generated in the laboratory are genetically identical to the parental strain. In our hands, the use of one primer, R108, is sufficiently discriminatory to distinguish between the isolates of different strains. - [Read Random Amplification of Polymorphic DNA (RAPD) Typing and Fingerprinting Protocol]

Category: Cytogenetics Protocols: Array CGH

Random Labelling DNA for array CGH Protocol. Sanger Institute - [Read Random Labelling DNA for array CGH Protocol]

Category: Nucleic Acid Modification Protocols: Random Primer Labeling Protocols

Protocol for random prime labeling of DNA. - [Read Random Prime Labeling of DNA Protocol]

Category: Nucleic Acid Modification Protocols: Random Primer Labeling Protocols

Protocol for random primer DNA labeling. - [Read Random Primer DNA Labeling Protocol]

Category: Nucleic Acid Modification Protocols: Random Primer Labeling Protocols

Protocol for random primer labeling of poly A and RNA. Includes: Random primer reaction; cDNA purification; Hybridization; Washing; Stripping of membranes. - [Read Random Primer Labeling of PolyA and RNA Protocol]

Category: Yeast Cell Culture Protocols: Yeast Sporulation Protocols

Random spore analysis is used to examine meiotic recombination without performing tetrad analysis. - [Read Random Spore Analysis in Yeast Protocol]

Category: DNA Protocols: cDNA Library Protocols

Protocol describes the generation of cDNA probes from poly(A)+ mRNA using random oligonucleotide primers. Probes of this type are used for differential screening of cDNA libraries. - [Read Synthesis of cDNA Probes from mRNA Using Random Oligonucleotide Primers Protocol]

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

Protocol describes the use of PCR to characterize a peptide library encoded in a plasmid vector. In this example, the library was obtained by transforming bacteria with the ligation reaction at the end of Use of PCR to Prepare a Double-Stranded DNA Library Encoding Random Peptides. - [Read Use of PCR for Quality Control of a Peptide DNA Library Protocol]

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

PCR is used as a preparative tool for the synthesis of a high-complexity double-stranded DNA library. In the example presented here, a mixture of synthetic oligonucleotides is used to synthesize a random peptide NNK library, where K is either T or G. The exclusion of A and C nucleotides at the third position decreases the occurrence of stop codons but still allows codons for all 20 amino acids. - [Read Use of PCR to Prepare a Double-Stranded DNA Library Encoding Random Peptides Protocol]