. 2014 Apr 11;14:90. doi: 10.1186/1471-2180-14-90

Table 3.

Primers used for targeted gene deletions

Primer name	Sequence 5′-3′	Purpose
pyrGN2	CACATGCCTCATTTTGACCA	Mutant confirmation
PyrtpsAup	ACCGTTGGAAGGTGGGATCCTATGGATCTCAGAA	Amplifies pyrG with 3' tpsA overhangs
PyrtpsAdown	CCTTTCAGAATGAGTGTGAGCGGATAACAATTTC	Amplifies pyrG with 3' tpsA overhangs
tpsAup	CCATCTGTCTAGCTCTTCATCCCC	tpsA, upstream fragment
tpsApyrup	GATCCATAGGATCCCACCTTCCAACGGTGTAGAGACTCC	tpsA, upstream fragment
tpsApyrdown	TTATCCGCTCACACTCATTCTGAAAGGTGGGGTTTTC	tpsA, downstream fragment
tpsAdown	GCAAGATTCCCGCATCCATC	tpsA, downstream fragment
tpsAupN1	CAACCCCACCAGTTCTCTCAAG	Amplification of KO-fragment
tpsAdownN1	AAAGGGAGTTCCAAGCAGCCTG	Amplification of KO-fragment
pyrtpsBup*	ATCTGCTCTGCCTGGGATCCTATGGATCTCAGAA	Amplifies pyrG with 3′ tpsB overhangs
pyrtpsBdown	CTGCCCATCACCATGTGAGCGGATAACAATTTC	Amplifies pyrG with 3′ tpsB overhangs
tpsBup*	TTGAACCCTTGAAACCGAACAC	tpsB, upstream fragment
tpsBpyrGup*	GATCCATAGGATCCCAGGCAGAGCAGATACTTACCCGTC	tpsB, upstream fragment
tpsBpyrGdown	TTATCCGCTCACATGGTGATGGGCAGACGATTG	tpsB, downstream fragment
tpsBdown	TGCTAAAGAGGGTGTGGGATTG	tpsB, downstream fragment
tpsBupN3	TCCCGATTGGTAGAATCCCTAAAG	Amplification of tpsB KO-fragment
tpsBdownN3	CATGCGAAAATGACAGGAACATTC	Amplification of tpsB KO-fragment
pyrGuphind	TAAAAGCTTCTATATTGATCCTTA	pyrG, KO of tpsC
pyrGdown	TGTGAGCGGATAACAATTTC	pyrG, KO of tpsC
tpsCupN-2	TGCCGAATTGACGTGCGTAGAG	Cloning of tpsC
tpsCdownN-2	TGGTGGTGAACCTTTCGTTGTTC	Cloning of tpsC
tpsCupN5	CCCTCCATACTTACTCCATACATCTCG	Amplification of tpsC KO-fragment
tpsCdownN5	CCAGCTTGACACATCCAACATAAC	Amplification of tpsC KO-fragment
pyrtppAup	CCTGTCCCCGCTTCAAGAAAGGGATCCTATGGATCTCAGAA	pyrG with 3′ tppA overhangs
pyrtppAdown	GAGTCATCAGTGCTGCTTTCTGCTGTGAGCGGATAACAATTTC	pyrG with 3′ tppA overhangs
TppAup	TGTTGGAAGCGTCTTTCTGCC	tppA, upstream fragment
tppApyrup	TTCTGAGATCCATAGGATCCCTTTCTTGAAGCGGGGACAGG	tppA, upstream fragment
tppApyrdown	GAAATTGTTATCCGCTCACAGCAGAAAGCAGCACTGATGACTC	tppA, downstream fragment
tppAdown	TGTCCGATTGGGGGTGATTG	tppA, downstream fragment
tppAupN1	TGAGGAGGCGTTGTCAAAAGATAG	Amplification of tppA KO-fragment
tppAdownN1	CGATTGGGGGTGATTGGCTTAC	Amplification of tppA KO-fragment
pyrtppBup	CGGTAGGTTAGGGATCCTATGGATCTCAGAA	Amplification of A. oryzae pyrG with 3′ tppB overhangs
pyrtppBdown	GTTTGTCTTGTGTGAGCGGATAACAATTTC	Amplification of A. oryzae pyrG with 3′ tppB overhangs
tppBup	ATACCAAGCAATCGCCCAAGCCAG	tppB, upstream fragment
tppBpyrGup	TCCATAGGATCCCTAACCTACCGCCCAAAGAGAGAGC	tppB, upstream fragment
tppBpyrGdown	TTGTTATCCGCTCACACAAGACAAACGATGCGGAATG	tppB, downstream fragment
tppBdown	CGTATCCTGGACTTTCAGCACG	tppB, downstream fragment
tppBupN1	TTTTCGACCTTGGTGGGTGCTTCC	Amplification of tppB KO-fragment
tppBdownN1	GAGACATTGTCGGTCAGTGAGGTAG	Amplification of tppB KO-fragment
pyrtppCup	TGTCCTTCAGGGATCCTATGGATCTCAGAA	pyrG with 3′ tppC overhangs
pyrtppCdown*	CTGTTCAGCATTGTGAGCGGATAACAATTTC	pyrG with 3′ tppC overhangs
tppCup	ATGAGGTGATAGTCGTGGACCCAG	tppC, upstream fragment
tppCpyrup	TCCATAGGATCCCTGAAGGACAAAGACAGGCTGAAG	tppC, upstream fragment
tppCpyrdown*	TTGTTATCCGCTCACAATGCTGAACAGATGATCCCCAG	tppC, downstream fragment
tppCdown*	TCGAGGTAGAGGTTCCCTTTCG	tppC, downstream fragment
tppCupN1*	CGATAGTCTTTGCGAACAGACGG	Amplification of tppC KO-fragment
tppCdownN1*	CGAGGTAGAGGTTCCCTTTCGATG	Amplification of tppC KO-fragment
tpsBupN1	CCCTTTCCCGATTGGTAGAATC	Amplification of tpsB/tppC double mutant

*Also used in the design of the tpsB/tppC double mutant.