The Polystichum Homepage

Welcome to the website for the hollyferns, members of the fern genus Polystichum and allies.
This website is maintained for the use of hollyfern enthusiasts and scholars
throughout the world by Dave Barrington at the University of Vermont in the United States.

Polystichum Homepage

Portrait of the Genus

Taxonomy

Species List

Diversity Map

Interactive Keys

Phylogeny

Cytology (PDF)

Barrington Web Page
(with publications)

Morphological Characters

Morphological Analysis

Lab Protocols

Nuclear Genes 1. Polystichum PGIc14-16 Protocol

Modified from Ishikawa et al., 2002

PRIMERS

LiPgiC 14F

GGGTCTTTTGAGTGTTTGG

LiPgiC 16R

GTTGTCCATTAGTTCCAGG

MIXTURE

Chemical Amount to add to 25uL reaction Concentration Final Concentration
PCR BUFFER 2.5uL 10X 1X
dNTPs 0.5uL 10mM 200uM
BSA 0.5uL 10mM 100ug/mL
ddH2O 17.875uL    
Taq 0.125uL 5U/uL 0.625U
primer F 1.25uL 10uM 1.0uM
primer R 1.25uL 10uM 1.0uM

PROGRAM

PCR cycles Time   Temperature°C
Initial denaturation 7 minutes @ 95°C
40 cycles X 30 seconds @ 94°C
  1 minute @ 56°C
  1 minute @ 72°C
Final extension 4 minutes @ 72°C

References

Ishikawa, H., Y. Watano, K. Kano, M. Ito, and S. Kurita. 2002. Development of primer sets for PCR amplification of the pgiC gene in ferns. Journal of Plant Research 115: 65-70.

Please be in touch with me with your questions about and insights into the holly ferns -