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Table 3 PCR analysis of hTERT ASPSs in lung and colon tumour/adjacent tissue cDNA samples

From: Characterization of novel alternative splicing sites in human telomerase reverse transcriptase (hTERT): analysis of expression and mutual correlation in mRNA isoforms from normal and tumour tissues

Tissue Sample 1) Ins-i2 [1–349] Ins-i2 [182–349] Ins-i2 [?–5273] Ins-i4 [1–38] Del-e6 [1–36] Del- e7e8 Del-e11 Ins-i14 [623–781] Ins-i14 [623–705] Ins-i14 [623–703] α-tub 2)
Lung A 25.2 0.0 *7.3 5.6 7.3 71.5 0.0 8.5 0.0 0.0 77.1
Adjacent B N N N 0.0 0.0 99.9 0.0 0.0 0.0 0.0 70.6
  C 0.0 0.0 0.0 0.0 0.0 0.0 0.0 47.5 0.0 0.0 61.9
  D N N N 0.0 0.0 *100.0 N 0.0 0.0 0.0 62.5
  E 0.0 0.0 0.0 100.0 100.0 100.0 N *51.3 0.0 0.0 61.1
Lung A 14.1 *0.5 13.3 1.8 2.9 39.5 0.0 1.4 0.4 *0.3 73.2
Tumour B 58.1 0.0 0.0 0.0 0.0 54.4 0.0 31.7 9.7 0.0 69.9
  C 10.7 0.0 9.1 5.4 *1.8 76.9 *12.7 10.6 *0.3 0.0 73.2
  D 0.0 0.0 0.0 0.0 0.0 76.7 0.0 18.5 0.0 0.0 79.3
  E 7.0 *0.1 *42.8 6.3 6.2 82.6 5.4 8.5 0.8 0.9 68.4
Colon A 0.0 0.0 0.0 *24.2 14.8 75.3 0.0 0.0 0.0 *2.9 53.7
Adjacent B 0.0 0.0 0.0 0.0 8.8 64.5 0.0 *3.6 0.0 0.0 64.4
  C 0.0 0.0 0.0 0.0 *4.7 65.7 0.0 10.2 0.0 0.0 55.9
  D 0.0 0.0 0.0 0.0 0.0 53.4 0.0 2.7 0.0 0.0 45.6
  E 0.0 0.0 0.0 0.0 0.0 81.0 0.0 *0.3 0.0 7.3 54.4
  F 0.0 *100.0 0.0 N 0.0 57.4 0.0 0.0 0.0 0.0 82.4
  G *0.1 *0.4 50.0 20.4 7.3 57.1 *0.6 3.1 0.0 0.0 88.9
Colon A 0.0 0.0 *100.0 0.0 0.0 69.7 0.0 0.0 0.0 0.0 57.3
Tumour B 29.2 0.0 0.0 19.6 22.3 63.2 0.0 16.6 0.0 *2.0 70.2
  C 0.0 0.0 0.0 0.0 0.0 76.6 0.0 40.2 0.0 0.0 67.8
  D 0.0 0.0 0.0 0.0 0.0 *54.1 0.0 13.2 0.0 0.0 68.3
  E *6.2 0.0 0.0 8.8 0.0 81.5 0.0 8.0 0.0 0.0 60.1
  F 2.5 0.0 20.2 22.7 7.1 58.1 0.4 3.9 0.7 0.7 91.7
  G 0.0 0.0 94.9 4.2 5.3 53.2 0.0 4.5 0.5 0.7 90.8
Pooled 3) Tissue            hTERT 4)sum total
PCR x35 Lung adjacent 25.1 0.0 0.0 11.1 17.9 61.9 0.0 13.6 0.0 0.0 1.0
  Lung tumour 11.9 0.0 8.8 3.3 3.2 56.3 1.9 5.5 0.6 0.2 14.2
  Colon adjacent 0.0 0.0 37.6 8.0 5.1 64.5 0.0 3.0 0.0 0.7 1.8
  Colon tumour 6.7 0.0 69.6 6.4 5.3 55.0 0.2 4.8 0.6 0.7 1.5
Extrapolated             
Δ 55/35 5)   0.55 - 2.22 1.71 2.02 1.11 5.29 0.76 1.03 1.06  
Ampl. rate 6)   0.970 - 1.041 1.027 1.036 1.005 1.087 0.986 1.002 1.003  
PCR start 7) Lung adjacent 72.0 - 0.0 4.3 5.2 51.8 0.0 22.0 0.0 0.0  
  Lung tumour 34.1 - 2.2 1.3 0.9 47.1 0.1 8.9 0.6 0.2  
  Colon adjacent 0.0 - 9.3 3.1 1.5 53.9 0.0 4.9 0.0 0.7  
  Colon tumour 19.2 - 17.2 2.5 1.5 46.1 0.0 7.8 0.5 0.6  
  1. 1) Values are percentage ASPS of the total hTERT PCR products produced in each respective PCR reaction. ASPS products detected after 55 cycles of PCR only are denoted with an asterisk; remaining values are from 35 cycles of PCR. N; no PCR products (including normal full-length) detected after 35 or 55 cycles of PCR. 2)α-tubulin values are percentage of primer incorporated in product after 35 cycles of PCR. 3) For calculation of pooled data the samples were normalized to α-tubulin levels and PCR products produced after 35 cycles of PCR were combined before calculation of percentages (group of samples treated as one sample). 4) Sum totals (relative values) of all hTERT PCR products produced for a given tissue calculated from normalized values. 5) Average change in percentages from 35 to 55 cycles of PCR computed from individual samples; the number is a factor where 1 means no change. 6) Amplification rate per cycle compared to normal full-length product calculated by taking the 20th root of Δ. 7) Estimation of ASPS percentages by extrapolation of data from 35 cycles of PCR using the formula: (ASPS %)/(Ampl. rate)35.