Abstract
Polymerase chain reaction (PCR) enables amplification of specific DNA fragments for the detection and tracking of Cryptosporidium spp. Newly obtained DNA are compared to an ever-growing database of Cryptosporidium sequences with uncertain or outdated metadata in primary public repositories (i.e., EMBL/DDBJ/GenBank). Here, we describe standard operating procedures to obtain DNA sequences from Cryptosporidium spp. marker genes. Small-subunit ribosomal RNA gene, large-subunit ribosomal RNA gene, and glycoprotein 60 are amplified using conventional PCR. Amplified and sequenced genes are compared to a reference library of up-to-date curated gene sequences to identify Cryptosporidium species and variants.
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