Nov 12, 2017

Public workspaceCryptococcus species Genotype and Serotype Analysis V.1

DOI
dx.doi.org/10.17504/protocols.io.kr2cv8e
  • Junjun Sang1,
  • Jiu Yi2,
  • Wei Fang2
  • 1Fuzhou General Hospital;
  • 2Changzheng Hospital
Junjun Sang
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DOI: dx.doi.org/10.17504/protocols.io.kr2cv8e
Protocol Citation: Junjun Sang, Jiu Yi, Wei Fang 2017. Cryptococcus species Genotype and Serotype Analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.kr2cv8e
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: November 12, 2017
Last Modified: February 10, 2018
Protocol Integer ID: 8730
Keywords: Genotype, Serotype Analysis
Materials
MATERIALS
Reagent Ethyl alcohol, Pure 200 proof, for molecular biology Merck MilliporeSigma (Sigma-Aldrich)Catalog #E7023
Reagent isopropyl alcohol
ReagentYPD BrothBio Basic Inc.Catalog #SD7022.SIZE.250g
Genomic DNA was extracted according to the procedures of MasterPure™ Yeast DNA Purification Kit.
Genomic DNA was extracted according to the procedures of MasterPure™ Yeast DNA Purification Kit.

The internal transcribed spacer (ITS) regions including the 5.8S rDNA clusters were amplified by PCR using the ITS4 and ITS5 sets (ITS4:5-TCCTCCGCTTATTGATATGC-3, ITS5: 5-GGAAGTAAAAGTCGTAACAAGG-3).
The internal transcribed spacer (ITS) regions including the 5.8S rDNA clusters were amplified by PCR using the ITS4 and ITS5 sets (ITS4:5-TCCTCCGCTTATTGATATGC-3, ITS5: 5-GGAAGTAAAAGTCGTAACAAGG-3).

Acquired sequences were submitted to GenBank and BLAST analysis to determine the identity of the isolates.
Acquired sequences were submitted to GenBank and BLAST analysis to determine the identity of the isolates.

PCR fingerprinting using the single primer, M13 (5-GAGGGTGGCGGTTCT-3), was used to identify the genotype and serotype of the C. neoformans isolates.
PCR fingerprinting using the single primer, M13 (5-GAGGGTGGCGGTTCT-3), was used to identify the genotype and serotype of the C. neoformans isolates.