Agilent Technologies E5500 Series User's Guide Page 3
- Page / 87
- Table of contents
- TROUBLESHOOTING
- BOOKMARKS
Rated. / 5. Based on customer reviews
Contents
3
Fragment Library Preparation Using the AB Library Builder
™
System: 5500 Series SOLiD
™
Systems User Guide
About This Guide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Safety information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
CHAPTER 1 About the Products . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Library preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Product information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Kit contents and storage conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
CHAPTER 2 Prepare to Build the Library . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Procedural guidelines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Quantitate the DNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Shear the DNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Set up the AB Library Builder
™
System for size-selected or express fragment library
preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
CHAPTER 3 Build the library . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Start the run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Set up for a new run . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
CHAPTER 4 Nick-Translate the Library with Optional Amplification . . . . . . . . 29
Workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Nick-translate the library . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
(Optional) Nick-translate and amplify the library . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Quantitate the DNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
Check the size distribution of the library . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
(Optional) Pool equal molar barcoded libraries of similar size . . . . . . . . . . . . . . . . . . . . . . . . . 34
CHAPTER 5 Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Instrument error codes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
- AB Library Builder 1
- Contents 3
- About This Guide 7
- About the Products 9
- How to use the 10
- Kit contents 11
- System: 5500 Series SOLiD 12
- Systems User Guide 12
- Chapter1 About the Products 12
- Workflow 13
- Procedural guidelines 13
- Shear the DNA 14
- Quantitate the DNA 14
- 5. Remove the sheared DNA: 16
- Set up the AB Library Builder 17
- System for size-selected or 17
- Insert or change 18
- Load the racks and 19
- Load the cartridge rack 20
- Insert the racks 23
- Insert the tip and tube rack 25
- Build the library 27
- Set up for a new run 28
- Optional Amplification 29
- Purify the nick 30
- Prepare the 31
- Troubleshooting 35
- Chapter5 Troubleshooting 36
- Instrument error codes 38
- Ordering Information 39
- Required equipment 42
- Optional equipment 44
- Replacement parts 44
- Required consumables 45
- Optional consumables 46
- Supplemental Procedures 47
- System operation 48
- Manage the run 50
- Size-select the 51
- 250-bp marker 54
- Collection wells 54
- Sample wells 54
- ND-1000 55
- Spectrophotometer 55
- 2. Open the NanoDrop 56
- Supplemental Background 59
- Information 59
- Genomic DNA Sheared DNA 60
- Library molecule ligated 60
- Sequencers User 62
- Guide (Part no. 4456991) 62
- Library Construction 63
- Oligonucleotide Sequences 63
- System Kit 73
- Safety 77
- Chemical waste safety 78
- Chemical waste 79
- Waste disposal 79
- Biological hazard safety 80
- Documentation and Support 81
- Glossary 83
- P1-T Adaptor 84
- Standard Adaptor 84
- 4460965A 87
- Headquarters 87
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