This documents applies to KAPA Library Amplification Kits (0795897808986001), KAPA Library Amplification Kits (PCR Mix only) (07958943001, This will give an approximate concentration of 20 pM. PN7960093001 (-20☌) **may be ordered separately KAPA Primer Mix, 1mL, PN7960093001 (-20☌) This ERCC analysis pipeline for absolute quantification of RNA-seq data is published alongside our 2020 BioProtocols paper. KAPA SYBR FAST qPCR Master Mix (2X) Kits are a ready-to-use cocktail containing all components (except primers Repeat steps 1 and 2 to produce 3 independent dilutions of the library template. Concentrations of six different diluted libraries were determined using either the sparQ Universal Library Quant Kit on Quantabio Q qPCR cycler or the Roche KAPA Library Quantification Kit on Bio-Rad CFX following the manufacturer’s recommended protocol.
Data analysis 7 Working example 8 Data Analysis and Interpretation 9. Dilution Factor Insert a numerical value determined by the dilution factor of the library material. MM9 genome files were used in our analysis for the paper. Results from sparQ and Roche KAPA Library Quant Kits were highly correlated. The KAPA SYBR FAST DNA Polymerase and proprietary buffer system improves the amplification efficiency of difficult targets, including both GC- and AT-rich templates.
(fluorescence), quantification cycle (Cq), linearity, and sensitivity. Template (DNA standard or library) 4 µL Table 2: Thermocycling parameters Step Temperature Duration Cycles Initial denaturation 95☌ 5 min 1 Denaturation 95☌ 30 sec Annealing/Extension 35 (data acquisition) 60☌ 45 sec Melt curve analysis 65☌ - 95☌ Methods KAPA Library Quantification Kit contains all reagents required You need a genome fasta file and a gtf file with gene features and coordinates. New Method from Acquired Scan Data… (with Library Search) Use Library Method to identify and create compounds.