Laboratory Protocols

Universidade Federal do Amazonas (UFAM), Departamento de Biologia, ICB, 69077-000 Manaus, AM, Brasil

Biology Department, University of Puerto Rico - Rio Piedras (UPR-RP), San Juan, PR, 00931, Puerto Rico

Microsatellite development protocol

We are currently using an enrichment protocol. In short, a repeat DNA of interest, a microsatellite, is bound to a iron metal bead via a biotin-streptavidin link. Total genomic DNA is hybridized to the conjunct, washed, and then enriched DNA is eluted. If things go well, you can develop microsats in about a week.

A protocol in English or in Portuguese

General Laboratory Protocols

High molecular DNA extraction protocol [PDF]

Basic phenol-chloroform extraction protocol [PDF]

High salt extraction protocol [PDF]

CTAB extraction protocol [PDF]

DNA extraction protocol from teeth [PDF]

PCR buffers [PDF]

Cresol Red PCR dye [PDF]

Basic microsatellite PCR protocol [PDF]

Fluorescent microsatellite PCR protocol [PDF]

Fluorescent microsatellite PCR protocol with dynamic labeling [PDF]

Basic mtDNA PCR protocol [PDF]

Nested nDNA PCR protocol [PDF]

Basic cycle sequencing PCR protocol [PDF]

Cycle sequencing clean-up protocol [PDF]

AFLP protocol

We are currently using an AFLP protocol to study genetic diversity in organisms for which we have no microsatellite markers. The protocol consists of digestion of total genomic DNA with two restriction enzymes, a six-cutter and a four-cutter, and ligation of appropriate linkers. The six cutter linker primer contains a sequence compatible with protocols for dynamic fluorescent labeling.

A protocol in English or in Portuguese