Using recombinant glycosylated and non-glycosylated irisin as positive controls, we found no immune-reactive bands of the expected size in any biological samples.
In all cases, therefore, we obtained bands of the expected sizes in one single analysis with the four samples pooled together using conditions that allow each band to be well resolved.
7304 simple sequence repeats (SSR) were identified from 6509 unigenes, a total of 9930 primer pairs were designed, 50 primer pairs were randomly selected to validate of the usefulness, and 24 (48%) primer pairs produced bands of the expected size.
Double digestions that yielded bands of the expected molecular weight, and gene sequence analyses confirmed the absence of mutations.
After PCR amplification, products were run on 1 2 % agarose gels and inspected for bands of the expected size.
After PCR amplification, products were run on 1 2 % agarose gels and examined for bands of the expected size to confirm the DNA extractions were successful.
The bands of the expected sizes were removed and purified with GFX PCR DNA and Gel Band Purification Kit (GE Healthcare).
Recombinant plasmid pAN7-1-Chi67-1 was digested witHindIIIII and verified by electrophoresis, and two bands of the expected size were observed (Fig. 1c).
PCR gave bands of the expected sizes ranging from 1.1 to 1.5 Kb.
The electrophoretic bands of the expected size were subcloned and sequenced.
If bands of the expected size appeared, we purified those products using a QIAQuick PCR purification kit.
