Table 3. Observed and Expected sequence distributions sorted by 5′ tag composition.

5′Tag	Sequences from forward primer	Sequences from reverse primer	Total sequences	Expected sequence frequency	Deviation
AA	141	115	256	274.75	−18.75
AC	179	113	292	274.75	17.25
AG	68	65	133	274.75	−141.75
AT	95	93	188	274.75	−86.75
CA	237	231	468	274.75	193.25
CC	305	326	631	274.75	356.25
CG	291	286	577	274.75	302.25
CT	263	258	521	274.75	246.25
GA	171	153	324	274.75	49.25
GC	114	93	207	274.75	−67.75
GG	166	146	312	274.75	37.25
GT	178	152	330	274.75	55.25
TA	127	86	213	366.33	−153.33
TC	113	111	224	366.33	−142.33
TG	105	87	192	366.33	−174.33
TT	110	100	210	366.33	−156.33
4A*	45 (gcta)	60 (gtca)	105	183,16	−78,16
4B*	48 (tcag)	50 (tgac)	98	274,75	−176,75
4C*	114 (ctag)	100 (tagc)	214	274,75	−60,75
Total	2870	2625	5495	5495

^*Sequence of the tetranucleotide tag in parentheses

Expected sequence frequencies are calculated to account for the number of initial PCRs commencing from each different 5′tag.