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Table 2 PCR cycling conditions and primer information used for double stranded cDNA generation and colony PCR analysis, as described in SMART cDNA Library Construction Kit [18].

From: Yeast functional screen to identify genetic determinants capable of conferring abiotic stress tolerance in Jatropha curcas

Process Cycling Conditions Primer information
cDNA synthesis 95°C - 2 min

95°C - 30 sec
65°C - 30 sec
68°C - 4 min
repeated 18-24 cycles

68°C - 15 min
Hold at 4°C
SMART IV™ Oligonucleotide (10 μM)
5'-AAGCAGTGGTATCAACGCAGAGTGGCCATTACGGCCGGG-3'
CDS III/3' PCR Primer (10 μM)
5'-ATTCTAGAGGCCGAGGCGGCCGACATG-d(T)30N-1N-3'
5' PCR Primer (10 μM)
5'AAGCAGTGGTATCAACG CAGAGT-3'
Colony PCR 95°C - 5 min

95°C - 30 sec
55°C - 30 sec
72°C - 3 min
repeated 29 cycles

72°C - 5 min
Hold at 4°C
Forward Primer: Gal 1
5'-AATATACCTCTATACTTTAACGTC-3'
Reverse Primer: V5/6XHIS
5'-ACCGAGGAGAGGGTTAGGGAT-3'