1005200_Sequencing_LTW_PairedEnd_GA2_v3 Sequencing LTF Paired End GA2 1005200 Rev B
Sequencing_LTF_Paire.. Sequencing_LTF_PairedEnd_GA2_1005200_RevB
User Manual: Sequencing LTF PairedEnd GA2 1005200 RevB
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Paired-End Sequencing on the Genome Analyzer II Lab Tracking Form 1 Flow cell ____________________ Run folder ____________________ Samples Loaded Lane 2 Sample ID Concentration (pM) 1 Sam ple 1 Con c. 1 2 Sam ple 2 Con c. 2 3 Sam ple 3 Con c. 3 4 Sam ple 4 Con c. 4 5 Sam ple 5 Con c. 5 6 Sam ple 6 Con c. 6 7 Sam ple 7 Con c. 7 8 Sam ple 8 Con c. 8 Cluster Generation Step Date Amplification Linearization, Blocking, and Primer Hybridization for Read 1 Paired-End Cluster Generation Kit v2 Lot Numbers Reagent Lot Number AMX1 AMX2 AT1 AT2 BMX EMX HP1 HP2 HP3 (Read 1) HP2 (Read 2) HT1 HT2 Part # 1005200 Rev. B Page 1 of 9 Paired-End Sequencing on the Genome Analyzer II Lab Tracking Form Reagent Lot Number LMX1 LMX2 RMX Reagent Volumes After Amplification Position Reagent 1 AMX1 3 HT2 9 AT1 12 HT1 Volume Reagent Volumes After Linearization, Blocking, and Primer Hybridization 3 Position Reagent 3 HT2 4 BMX 7 HP1 12 HT1 14 LMX1 17 HP3 Volume Preparing Sequencing Reagents for Read 1 SBS Sequencing Kit v3 Lot Numbers Reagent Lot Number IMX FFN SDP CLM SMX PR1 PR2 PR3 PW1 Part # 1005200 Rev. B Page 2 of 9 Paired-End Sequencing on the Genome Analyzer II Lab Tracking Form 4 Pre-Run Wash Expected Volume Delivered Volume 12 ml 5 Loading and Priming Reagents Weights of Material Delivered Position Reagent Weight Before Run Weight After Run 1 IMX 0.00 2 PW1 0.00 3 SMX 0.00 4 PR1 0.00 5 PR2 0.00 6 CLM 0.00 7 PR3 0.00 Priming Reagents Expected Volume Delivered Volume 6.4 ml 6 % Difference -100.00 Checking for Leaks and Reagent Delivery Port Position 5 Expected Volume: 800 μl Measured Volume 1: ____________________ Measured Volume 2: ____________________ Measured Volume 3: ____________________ % Difference Between Expected Volume and Measured Volume 3: -100 ____________________ Part # 1005200 Rev. B Page 3 of 9 Delivered Weight Paired-End Sequencing on the Genome Analyzer II Lab Tracking Form 7 Setting Best Focus for Channel A, Read 1 Z Position FQ Value 1 A300 1 C300 2 A200 2 C200 3 A100 3 C100 4 A0 4 C0 5 A-100 5 C-100 6 A-200 6 C-200 7 A-300 7 C-300 Position of Z for Lane 4: 8 _____________________ Adjusting Focus FQ Calculation Z Position A C G T Total A70 0 C70 0 G70 0 T70 0 TOT7 00 A60 0 C60 0 G60 0 T60 0 TOT6 00 A50 0 C50 0 G50 0 T50 0 TOT5 00 A40 0 C40 0 G40 0 T40 0 TOT4 00 300 A30 0 C30 0 G30 0 T30 0 TOT3 00 200 A20 0 C20 0 G20 0 T20 0 TOT2 00 100 A10 0 C10 0 G10 0 T10 0 TOT1 00 0 A0 C0 G0 T0 TOT0 -100 A100 C100 G100 T100 TOT100 -200 A200 C200 G200 T200 TOT200 -300 A300 C300 G300 T300 TOT300 A400 C400 G400 T400 TOT400 A500 C500 G500 T500 TOT500 A600 C600 G600 T600 TOT600 A700 C700 G700 T700 TOT700 Part # 1005200 Rev. B Page 4 of 9 Paired-End Sequencing on the Genome Analyzer II Lab Tracking Form 9 Performing Post-Run Procedures for Read 1 Run Fluid Delivery Data Lane Weight of Empty Tube Weight of Tube After Run Weight of Fluid Delivered During Run 0.00 0.00 1 2 4 0.00 0.00 5 0.00 6 0.00 7 0.00 8 0.00 3 10 Step Read 1 Data Transfer Description Date Scan before Scan after 1a Images 1b Focus images (if stored) 1c Log files IB before IB after 1d Configuration files HSB before HSB after 1e Calibration files CM before CM after 3 Delete Run Folder CB before CB after 11 IM after Loading and Priming Reagents for Read 2 Preparation Weights of Material Delivered Position 10 11 12 13 15 16 Reagent Weight Before Run (g) Weight After Run (g) Delivered Weight (g) RMX IM before IM after IM deliv. LMX2 IB before IB after IB deliv. BMX HSB before HSB after HSB deliv. AMX2 CM before CM after CM deliv. AT2 Scan before Scan after Scan deliv. HP2 CM before IM after IM deliv. Part # 1005200 Rev. B Page 5 of 9 0 0 0 0 0 0 Paired-End Sequencing on the Genome Analyzer II Lab Tracking Form Position Reagent 19 21 Weight Before Run (g) HP3 CM before HT2 Water before CM after 1 Water after 8 Weight After Run (g) Delivered Weight (g) 2 CM deliv. -1 3 Water deliv. 5 Reagent Volumes After Priming and Read 2 Preparation Position Reagent 10 RMX 11 LMX2 12 BMX 13 AMX2 15 AT2 16 HP2 19 HP3 21 HT2 12 Volume After Priming Preparing Sequencing Reagents for Read 2 SBS Sequencing Kit v3 Lot Numbers Reagents Lot Number IMX FFN SDP CLM SMX PR1 PR2 PR3 PW1 Part # 1005200 Rev. B Page 6 of 9 Volume After Read 2 Preparation Paired-End Sequencing on the Genome Analyzer II Lab Tracking Form 13 Loading Sequencing Reagents for Read 2 Weights of Material Delivered Position Solution 1 IMX 0.00 2 PW1 3 SMX 0.00 0.00 4 PR1 0.00 5 PR2 0.00 6 CLM 0.00 7 PR3 0.00 14 Weight before Run Weight after Run Checking for Leaks and Reagent Delivery Port Position 5 Expected Volume: 800 μl Measured Volume 1: ____________________ Measured Volume 2: ____________________ Measured Volume 3: ____________________ % Difference Between Expected Volume and Measured Volume 3: 15 -100 ____________________ Setting Best Focus for Channel A, Read 2 Z Position FQ value 1 A300 1 C300 2 A200 2 C200 3 A100 3 C100 4 A0 4 C0 5 A-100 5 C-100 6 A-200 6 C-200 7 A-300 7 C-300 Position of Z for Lane 4: Part # 1005200 Rev. B _____________________ Page 7 of 9 Delivered Weight Paired-End Sequencing on the Genome Analyzer II Lab Tracking Form 16 Adjusting Focus for Read 2 FQ Calculation Z Position A C G T Total A70 0 C70 0 G70 0 T70 0 TOT7 00 A60 0 C60 0 G60 0 T60 0 TOT6 00 A50 0 C50 0 G50 0 T50 0 TOT5 00 A40 0 C40 0 G40 0 T40 0 TOT4 00 300 A30 0 C30 0 G30 0 T30 0 TOT3 00 200 A20 0 C20 0 G20 0 T20 0 TOT2 00 100 A10 0 C10 0 G10 0 T10 0 TOT1 00 0 A0 C0 G0 T0 TOT0 -100 A100 C100 G100 T100 TOT100 -200 A200 C200 G200 T200 TOT200 -300 A300 C300 G300 T300 TOT300 A400 C400 G400 T400 TOT400 A500 C500 G500 T500 TOT500 A600 C600 G600 T600 TOT600 A700 C700 G700 T700 TOT700 17 Performing Post-Run Procedures for Read 2 Run Fluid Delivery Data Lane Weight of Empty Tube Weight of Tube after Run Weight of Fluid Delivered during Run 1 0.00 2 0.00 3 4 0.00 0.00 5 0.00 6 0.00 7 0.00 8 0.00 Part # 1005200 Rev. B Page 8 of 9 Paired-End Sequencing on the Genome Analyzer II Lab Tracking Form Post-Run Wash Weight of Tube after Wash: ____________________ Weight of Empty Tube: ____________________ Weight of Fluid Delivered: 0 ____________________ 18 Sign-off Date: ____________________ Name: ____________________ Signature: ____________________ Part # 1005200 Rev. B Page 9 of 9
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