42 u00b0 c 5 ul samples of each reaction mix were subjected to slot blot analysis and using radiolabelled bb10 4 oligonucleotide as a probe for the presence of 3sr reaction products in column 1 bb113 was used as the 5 39 primer which was paired with either bb112 h16 686 h16 743 or h16 773 the results indicate that bb112 and bb773 yield significant signals above the background sample in slot 1 this slot has a sample of a 3sr reaction run without added template in the second column a similar analysis was performed using primer h16 58 as a 5 39 primer in this case primer h16 743 yields the most significant signal above background these results demonstrate the importance of primer selection since this example shows not every 5 39 primer can support amplification with every 3 39 primer table 1__________________________________________________________________________oligonucleotides used in 3sr in situ amplification__________________________________________________________________________designator gene sequence__________________________________________________________________________ bb111 e7 atg cat gga gat aca cct aca ttgseq id no 2 bb112 e7 gca caa ccg aag cgt aga gtc acaseq id no 3 bb113 e6 atg cag caa aag aga act gca atgseq id no 4 bb114 e6 cag ctg ggt ttc tct acg tgt tctseq id no 5h16 58 e6 gga ccc aca gga gcg acc cag aaa gtseq id no 6 h16 773 e7 tct gag aac aga tgg ggc aca caaseq id no 7 h16 743 e7 gtg tgc cca tta aca ggt ctt ccaseq id no 8 h16 686 e7 a caa ccg aag cgt aga gtc aca ctseq id no 9__________________________________________________________________________detector oligonucleotides sequence__________________________________________________________________________bb4 e6 taa ctt tct ggg tcg ctc ctg tggseq id no 10 gtc ctgbb109 e7 ata gat ggt cca gct gga caa gcaseq id no 11 gaa.