Directed Electronics equalizer 2500 Betriebsanweisung PDF herunterladen (Seite 37)

Load and Prime Reagents

HiSeq 2500 System User Guide

7 Place each reagent bottle onto the rack in the associated numbered position. Make sure

that the conical end of the bottle rests in the indentation on the base of the rack.

Position

Reagent Description

IRM Incorporation Reagent Master Mix

PW1 25 ml of PW1 or laboratory-grade water

USM Universal Scan Mix

SBS Buffer 1 (SB1) High Salt Buffer

SBS Buffer 2 (SB2) Incorporation Wash Buffer

CRM Cleavage Reagent Mix

SBS Buffer 3 (SB3) Cleavage Buffer

Table 1 Reagent Positions

8 Add 25 ml of PW1 or laboratory-grade water to the bottle in position 2.

9 Slide the reagent rack into the reagent compartment, aligning the rack with the raised

guide on the floor of the compartment.

10 Lower the sippers into the sequencing reagent bottles as follows:

a Pull the sipper handle towards you and then lower the sipper handle.

b Visually inspect the sippers to make sure that they do not bend as they lower into

the funnel caps.

c Release the sipper handle into the slot on the bottom end of the groove.

11 Select the PW1 (25 ml) loaded checkbox.

Load Indexing Reagents

1 Record the weight of each reagent on the lab tracking form.

2 Make sure that the paired-end rack is not in use on the adjacent flow cell. Steps that

use the paired-end rack include Read2 resynthesis, Index 1 (i7) Read preparation, and

Index 2 (i5) Read preparation.

3 Raise the sippers for the paired-end reagent rack using the following motion:

a Pull the handle towards you and then raise the handle.

b Release the handle into the slot on the top end of the groove. Make sure that the

handle rests securely in the slot.

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Directed Electronics equalizer 2500 Betriebsanweisung Seite 37