Agilent Technologies E5500 Series User's Guide download pdf (Page 54)

Fragment Library Preparation Using the AB Library Builder

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System: 5500 Series SOLiD

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Systems User Guide

AppendixB Supplemental Procedures

(Optional) Size-select and pool libraries

3. When the 250-bp band (~240–270-bp region) from the marker (ladder) lane is at

the top of the collection well, stop the run if the run has not already stopped:

Note: After amplification, the total size of the product is ~240–270 bp, and the

estimated insert size after size selection is ~150–180 bp.

Collect the sample from the SOLiD

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Library Size Selection Gel

Collect the solution from the wells and pool according to samples.

2. Wash each collection well with 25 µL with Nuclease-free Water, then retrieve the

wash solution with the solution collected in Step 6.

3. (Optional) Concentrate the DNA with a SOLiD

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Library purification column.

(Op tional ) Pool

remaining libraries

that will be

combined into a

single emulsion

1. Quantitate the libraries to be pooled by qPCR (see “Quantitate the DNA” on page

33).

2. Mix together equal molar amounts of each barcoded library in an appropriately

sized LoBind Tube. Vortex the tube.

STOPPING POINT Store the purified DNA in Elution Buffer (E1) at – 20°C, or proceed

directly to emulsion PCR, as describe in the SOLiD

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EZ Bead

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Emulsifier Getting

Started Guide (Part no. 4441486).

250-bp marker

Collection wells

Sample wells

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Agilent Technologies E5500 Series User's Guide Page 54