PROCEED AMP 5 User Manual download pdf (Page 21)

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 frozen, allow

them to thaw to room temperature, and then gently invert the reagent tubes at least 10

times to mix contents.

} Ensure the water circulator is filled to the appropriate level.

} Turn on the water circulator. Set it to a temperature that brings the Chamber Rack to 44°C

at equilibrium.

} Remove bubbles trapped in the Chamber Rack.

} Test several locations on the Chamber Rack, using the Illumina Temperature Probe. All

locations should be at 44°C ± 0.5°C.

Single-Base Extension

CAUTION

The remaining steps must be performed without interruption.

NOTE

If you are processing more than 8 BeadChips complete the reagent dispensing

step for each reagent for the first set of 8 BeadChips, then continue the same

reagent dispensing steps for the second set of 8 BeadChips. Then move to the last

set of 8 BeadChips before you start the incubation time.

Steps marked with an asterisk (*) indicate when you should follow this reagent

dispensing method.

[_] 1 When the Chamber Rack reaches 44°C, quickly place each Flow-Through Chamber

assembly into the Chamber Rack.

For 4 BeadChips, place the Flow-Through Chambers in every other position, starting at 1,

in the first row of the Chamber Rack. For larger numbers of BeadChips, fill all positions

in the first row, then the second and third.

[_] 2 Into the reservoir of each Flow-Through Chamber, dispense:

[_] a 150μl RA1. Incubate for 30 seconds. Repeat 5 times.

[_] 1 [_] 2 [_] 3 [_] 4 [_] 5 [_] 6

[_] b 225μl LX1. Repeat once*. Incubate for 10 minutes.

[_] 1 [_] 2

[_] c 225μl LX2. Repeat once*. Incubate for 10 minutes.

[_] 1 [_] 2

[_] d 300μl EML. Incubate for 15 minutes.

[_] e 250μl 95% formamide/1 mM EDTA. Incubate for 1 minute. Repeat twice.

[_] 1 [_] 2 [_] 3

[_] f Incubate 5 minutes.

[_] g Begin ramping the Chamber Rack temperature to the temperature indicated on the

SML tube.

[_] h 250μl XC3. Incubate for 1 minute. Repeat twice*.

[_] 1 [_] 2 [_] 3

[_] 3 Wait until the Chamber Rack reaches the correct temperature.

Illumina Infinium LCG Quad Assay, Manual Protocol

Experienced User Card

Part # 15025909 Rev. A

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