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A set of three "housekeeping" gene primer pairs (Sigma) was used to calibrate template amount (Table 2).
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Beta-actin was used as an internal control to calibrate the cDNA template for all samples.
For relative quantification, two house-keeping genes were used as calibrators for each alga in this study to verify successful reverse transcription and to calibrate the cDNA template.
Two clam β-actin primers, P7 and P8 (Table 2) were used to amplify a 121 bp fragment as internal control to verify the successful reverse transcription and to calibrate the cDNA template.
Two β-actin primers, AF and AR (Table 1), were used to amplify a 94-bp fragment as an internal control to verify the successful transcription and to calibrate the cDNA template for corresponding scallop samples.
A template was calibrated for each shoulder at the beginning of the first session.
To calibrate the reaction, the cDNA template was serially diluted and amplified each time along with every gene targets.
The number of bands at a length below 1,500 bp were counted, calibrated in regard to PCR template concentration and presented as the number of telomeres below a length of 1,500 bp per genome equivalent of template DNA.
All lengths were measured in mm in a standardized fashion and calibrated using a 10-cm sized template for elimination of magnification errors.
For accurate pitch targeting, the template attached on the tablet is carefully calibrated.
A template was photographed alongside the target ulcer to calibrate the image in the image analysis software (see supplementary figure).
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