### II. Calculating Expression Values
-To prepare the reference sequences, you should run the
+To calculate expression values, you should run the
'rsem-calculate-expression' program. Run
rsem-calculate-expression --help
to get usage information or visit the [rsem-calculate-expression
documentation page](rsem-calculate-expression.html).
-Note: RSEM no longer provides nu values. Instead, RSEM provides
-nrf(normalized read fraction), which is a normalized version of theta
-vector excluding theta_0.
-
#### Calculating expression values from single-end data
For single-end models, users have the option of providing a fragment
=item B<input>
-SAM/BAM formatted input file. If "-" is specified for the filename, SAM/BAM input is instead assumed to come from standard input. SAM/BAM format used is SAM Spec v1.2. RSEM requires all alignments of the same read group together. For paired-end reads, RSEM also requires the two mates of any alignment be adjacent. See Description section for how to make input file obey RSEM's requirements.
+SAM/BAM formatted input file. If "-" is specified for the filename, SAM/BAM input is instead assumed to come from standard input. RSEM requires all alignments of the same read group together. For paired-end reads, RSEM also requires the two mates of any alignment be adjacent. See Description section for how to make input file obey RSEM's requirements.
=item B<reference_name>
sort -k 1,1 -s input.sam > input.sorted.sam
-The SAM/BAM format RSEM uses is v1.2.
+The SAM/BAM format RSEM uses is v1.3. However, it is compatible with old SAM/BAM format.
The user must run 'rsem-prepare-reference' with the appropriate reference before using this program.