Research Article

Comparative Analysis of the Methanogen Diversity in Horse and Pony by Using mcrA Gene and Archaeal 16S rRNA Gene Clone Libraries

Table 1

PCR primers used in real-time PCR assays and in clone library analysis.

Application TargetName (direction)Sequence (5′→3′)Annealing temperature (°C)Number of cyclesProduct size (bp)Reference

Quantitative real-time PCRAnaerobic fungi q-pcr (f)GAGGAAGTAAAAGTCGTAACAAGGTTTC  60 40 120 [29]
q-pcr (r) CAAATTCACAAAGGGTAGGATGATT
Methanogen q-mcra (f)TTCGGTGGATCDCARAGRGC  60 40 141 [23]
q-mcra (r)GBARGTCGWAWCCGTAGAATCC
Total bacteria1114 (f)CGGCAACGAGCGCAACCC  60 40 130 [29]
1275 (r)CCATTGTAGCAGGTG

16S rRNA libraryMethanogenMet 86 (f)GCTCAGTAACACGTGG  53 30 1254 [30]
Met 1340 (r)CGGTGTGTGCAAGGAG

mcrA libraryMethanogenmcrA(f)GGTGGTGTMGGATTCACACARTAYGCWACAGC  58 30 480 [31]
mcrA (r)TTCATTGCRTAGTTWGGRTAGTT

f: forward; r: reverse.