Primers used for PCR and sequencing are listed. For amplification of 23S ribosomal gene
DNA,New100R2was used in combination of either of New061F2,New022F orNew041R2.
New100R2 is a sequence of 16S ribosomal genes which is located upstream of 22S
ribosomal gene, New061F2, New022F and New041R2 are the sequences of downstream
regions of 23S ribosomal genes. These primer pairs made it possible to amplify DNA of
about 6 kb which covers the whole gene of the respective copy of three 23S ribosomal
genes. Primers 0770FwrdHindIII and 0770RevXhoI were used to amplify 1 kb DNA which
contains the entire rluD gene and BfmS12F and BfmS12Rwere used for amplification of the
rpsL gene of 400 bp. All primers listed here were used for the sequencing of a respective
target gene, and designed by us on the basis of the whole genome sequence of B. bifidum
YIT 4007