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Reference Annotation

Parameter File

Reference Genome 

Parameter

Expression
NB_MOLECULES5,000,000Number of RNA molecules initially in the experiment
TSS_MEAN25Average deviation from the annotated transcription start site (TSS)
POLYA_SCALE300Scale of the Weibull distribution, shifts the average length of poly-A tail sizes
POLYA_SHAPE2Shape of the Weibull distribution describing poly-A tail sizes
Fragmentation
FRAG_SUBSTRATERNASpecifies RNA as the substrate of fragmentation
FRAG_METHODURUniform random fragmentation
FRAG_UR_ETA170

Average expected framgent size after fragmentations, i.e., number of breaks per unit length (exhautiveness of fragmentation)

FRAG_UR_D01Minimum length of fragments produced by UR fragmentation
FRAG_UR_DELTANaNGeometry of molecules in the UR process depends logarithmically on molecule length
Reverse Transcription
RTRANSCRIPTIONYESSwitch on the reverse transcription
RT_PRIMERRHUse random hexamer primers used for first strand synthesis
RT_MOTIFdefault

A default PWM of the current Illumina protocol is used

RT_LOSSLESSYESFlag to force every molecule to be reversely transcribed
RT_MIN500Minimum length observed after reverse transcription of full-length transcripts
RT_MAX5,500Maximum length observed after reverse transcription of full-length transcripts
Amplification and Size Segregation
PCR_DISTRIBUTIONdefaultDefault PCR distribution with 15 rounds and 20 bins
GC_MEAN0.5Mean value of a gaussian distribution that reflects GC bias amplification probability
GC_SD0.1Standard deviation of a gaussian distribution that reflects GC bias amplification probability
FILTERINGYESEnables size filtering of fragments

SIZE_DISTRIBUTION

nullEmploy an empirical Illumina fragment size distribution
SIZE_SAMPLINGMHThe Metropolis-Hastings algorithm is used for filtering
Sequencing
READ_NUMBER15,000,000Produce 15 million reads
READ_LENGTH75Each read sequence is 75nt long
PAIRED_ENDYESPaired-end reads are simulated, two per fragment

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