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Cell Applications Inc. guarantees to provide the highest quality RNA available. A variety of Total
RNA's are prepared from primary cells and tissues from human and animal using a modified guanidium thiocyanate method. Purified RNA is ready for use in various downstream applications, including microarray
hybridization, PCR cDNA synthesis and RT-PCR analysis.
Stringent quality control tests were performed on each preparation to confirm the integrity and
stability of RNA.
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Distinct 28S and 18S ribosomal RNA were shown
on 1% denaturing gel before and after 37oC incubation.
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No visible Genomic DNA band of 23 kb was
detected.
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No genomic DNA contamination was detected
following PCR amplification using house-keeping gene G3PDH primers.
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A 2.2 kb product from mRNA was detected
following RT-PCR amplification using human heat shock gene Hsp60 primers.
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A discrete band of 2 kb was detected upon
hybridization with a human beta-actin cDNa probe in Northern Blot analysis.
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Primers were designed to detect mRNA for genes
of interest to confirm the phenotype of the cells.
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Fig 1.
Two distinct 28S and 18S ribosomal RNA bands were showed on 1% denatured agarose gel. Intensity of 28S is two times as that of 18S band with no degrading. Lane 1-4: Total RNA of RAOSMC, HFDPC, HUVEC, and HOb.
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Fig 2. RT-PCR to verify the
integrity of RNA and phenotype. Lane 1-4: Total RNA of RAOSMC, HFDPC, HUVEC, and HOb with HSP60 primers. Lane 5: Total RNA of stimulated human osteoblasts with osteocalin primers. Lane 6: RT product of re-differeciated human
chondrocytes with aggrecan primers.
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Fig 3.
The absence of contaminating genomic DNA was confirmed by PCR with G3PDH primers. Size of 450 bps target band was not seen on 1% agarose gel. Lane 1-4: Total RNA of RAOSMC, HFDPC, HUVEC and HOb. Lane 5: positive control of G3PDH cDNA.
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Fig 4. Human Total RNA was denatured and separated on 1% of denaturing agarose gel after which the RNA was transferred onto nylon membrane
and hybridized with human beta-actin probe. Lane 1: human pancreas total RNA. Lane 2: human lung Total RNA.
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