Fluidigm Access Array - Prepare the 20 X Primer Solutions

Access Array System for the Ion Torrent PGM Sequencing System: User Guide
Chapter5:Bidirectional Amplicon Tagging on the LP 48.48 IFC
Prepare the 20X Primer Solutions
28
Prepare the 20X Primer Solutions
NOTE If you are using primers provided by the Fluidigm Assay Design Group, refer to the
order informatics packet for correct primer concentrations.
1 Prepare the 20X primer solutions for 48 individual primer pairs as shown in the table
below. These will be loaded into the primer inlets of an IFC.
IMPORTANT Warm the 20X Access Array Loading Reagent to room temperature before
use.
Table 7. Example for ASY-AA product (Fluidigm): Dilution of target-specific primers
2 Vortex the 20X primer solutions for a minimum of 20 seconds and centrifuge for
30 seconds.
NOTE The final tagged TS forward and reverse primer concentrations are 4 M in the
20X primer solution. The final TS forward and reverse primer concentrations in the
Access Array reaction chamber are 200 nM.
123
456
789
10 11 12
13 14 15
16 17 18
19 20 21
22 23 24
25 26 27
28 29 30
31 32 33
34 35 36
37 38 39
40 41 42
43 44 45
46 47 48
123
456
789
10 11 12
13 14 15
16 17 18
19 20 21
22 23 24
25 26 27
28 29 30
31 32 33
34 35 36
37 38 39
40 41 42
43 44 45
46 47 48
PRIMER INLETS
H1
H3
SAMPLE INLETS
H2
H4
INTERFACE ACCUMULATOR
CONTAINMENT ACCUMULATOR
Component Volume
(μL)
Final
Concentration*
* The final concentration in this table refers to the amount of each listed component in 5 L of the final primer
mix.
50 M CS1-tagged TS forward primer/50 M CS2-tagged TS
reverse primer
8.0 4 M
20X Access Array Loading Reagent (Fluidigm) 5.0 1X
PCR Certified Water (Teknova) 87.0
Total 100