PROCEED AMP 5 User Manual

ILLUMINA PROPRIETARYCatalog # WG-901-5002Part # 15025909 Rev. AMay 2011Page 1 of 26Illumina Infinium LCG Quad Assay, Manual ProtocolExperienced User C

Resuspend the AMP5 Plate (Post-AMP)Page 10 of 26Part # 15025909 Rev. A• If you do not plan to proceed to the next step immediately, store the sealed A

Hybridize Multi BeadChip (Post-AMP)Hybridize Multi BeadChip (Post-AMP)Dispense the fragmented, resuspended DNA samples onto BeadChips. Incubate theBea

Hybridize Multi BeadChip (Post-AMP)Page 12 of 26Part # 15025909 Rev. ANOTEYou can also prepare the Hyb Chambers later, during the 30-minute cool down.

Hybridize Multi BeadChip (Post-AMP)[_] 4 On the lab tracking form, record the BeadChip barcode for each group of samples.[_] 5 Visually inspect all se

Hybridize Multi BeadChip (Post-AMP)Page 14 of 26Part # 15025909 Rev. A[_] 4 Place the Hyb Chamber in the 48°C Illumina Hybridization Oven so that the

Wash the BeadChip (Post-AMP)Wash the BeadChip (Post-AMP)Prepare the BeadChips for the staining process.Estimated Time• 20 minutes for 4 BeadChips• 30

Wash the BeadChip (Post-AMP)Page 16 of 26Part # 15025909 Rev. A[_] a Using powder-free gloved hands, hold the BeadChip securely and by the edges inone

Wash the BeadChip (Post-AMP)[_] 1 If you have not done so, fill the Multi-sample BeadChip Alignment Fixture with 150 mlPB1.If more than four BeadChips

Wash the BeadChip (Post-AMP)Page 18 of 26Part # 15025909 Rev. ACAUTIONPlace all assembled Flow-Through Chambers on the lab bench in a horizontalpositi

Single-Base Extension and Stain LCG BeadChip (Post-AMP)Single-Base Extension and Stain LCG BeadChip (Post-AMP)In this process, you use RA1 reagent to

Page 2 of 26Part # 15025909 Rev. AIllumina Infinium LCG Quad Assay, Manual ProtocolExperienced User Card

Single-Base Extension and Stain LCG BeadChip (Post-AMP)Page 20 of 26Part # 15025909 Rev. AConsumablesItem Quantity Storage Supplied ByRA1 10ml for 1-

Single-Base Extension and Stain LCG BeadChip (Post-AMP)Preparation} Place all reagent tubes in a rack in the order in which they will be used. If froz

Single-Base Extension and Stain LCG BeadChip (Post-AMP)Page 22 of 26Part # 15025909 Rev. AStain BeadChipNOTEIf you are processing more than 8 BeadChip

Single-Base Extension and Stain LCG BeadChip (Post-AMP)[_] 6 Shake the XC4 bottle vigorously to ensure complete resuspension. If necessary, vortexunti

Single-Base Extension and Stain LCG BeadChip (Post-AMP)Page 24 of 26Part # 15025909 Rev. AIllumina Infinium LCG Quad Assay, Manual ProtocolExperienced

Image BeadChip (Post-AMP)Image BeadChip (Post-AMP)Follow the instructions in the iScan System User Guide or HiScanSQ System User Guide to scanyour Bea

Image BeadChip (Post-AMP)Page 26 of 26Part # 15025909 Rev. AIllumina Infinium LCG Quad Assay, Manual ProtocolExperienced User Card

Make the AMP5 Plate (Pre-AMP)Make the AMP5 Plate (Pre-AMP)This process creates a AMP5 plate for DNA amplification. The DNA sample is denaturedwith 0.1

Make the AMP5 Plate (Pre-AMP)Page 4 of 26Part # 15025909 Rev. A• Columns 1, 3, 5, 7, 9, and 11 (48 samples)[_] 6 On the lab tracking form, record the

Fragment the AMP5 Plate (Post-AMP)Fragment the AMP5 Plate (Post-AMP)This process enzymatically fragments the amplified DNA samples. An end-pointfragme

Fragment the AMP5 Plate (Post-AMP)Page 6 of 26Part # 15025909 Rev. A[_] 4 Seal the AMP5 plate with the 96-well cap mat.[_] 5 Vortex the plate at 1600

Precipitate the AMP5 Plate (Post-AMP)Precipitate the AMP5 Plate (Post-AMP)Add PA1 and 2-propanol to the AMP5 plate to precipitate the DNA samples.Esti

Precipitate the AMP5 Plate (Post-AMP)Page 8 of 26Part # 15025909 Rev. A[_] 12 Over an absorbent pad, decant the supernatant by quickly inverting the A

Resuspend the AMP5 Plate (Post-AMP)Resuspend the AMP5 Plate (Post-AMP)Add RA1 to the AMP5 plate to resuspend the precipitated DNA samples.Estimated Ti