PCR QC
Obtaining what appears to be good high molecular weight genomic is only the first line
of QC for this protocol. The ultimate test is to see if the DNA can be used to amplify a
Genomic
DNA PCR product. The test case used in developing this protocol was leaf tissue from the
coleus plant (Coleus blumei). In order to test the DNA for PCR amplification the gene
encoding tyrosine aminotransferase (GenBank #AJ458993) was submitted for PCR
primer selection using the IDT SciTools software PrimerQuest. The software chose the
following sequences:
Tat FOR:5’- ATA AAC CCT GGG AAC CCA TGT GGA –3’
Tat REV: 5’- AAC TTT GGG CTC ATC AAA GTG CCG –3’
These sequences were synthesized and a PCR amplification carried out using the
conditions; 5 min. 94o
C; 35 x (30 sec. 94o
C, 30 sec. 57o
C; 30 sec 72o
C); 7 min. 72o
C.
Results of this amplification are shown below.