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MicroRNA profiling in Acute Lymphoblastic Leukemia (ALL)

ABSTRACT: MicroRNA (667) profiling was done using TaqMan MicroRNA Arrays, which contains megaplex primer Pools covering Sanger miRBase version10 of Taqman Low Density Arrays (TLDA) platform across 50 samples along with the normals consisting of T cell lineage Acute Lymphoblastic Leukemia (T-ALL) and B cell lineage Acute Lymphoblastic Leukemia (B-ALL) subtypes which consists peripheral blood (PB) and bone marrow (BM) samples. The above isolated RNA which displayed good RIN value, linearity (R2>0.96), were used for reverse transcription (RT) reactions with the help of TaqMan MicroRNA Reverse Transcription Kit followed by PCR Reaction (ABI 7900 HT) as per illustration provided in kit. Real-time PCR was performed using an Applied Biosystems 7900 HT- TLDA Real-Time PCR System. A pre -amplification step of cDNA with pre-amp megaplex pool primers was done to significantly enhance the ability to detect highly down regulated miRNAs. The TaqMan human microRNA arrays consists of two plates, pool A and pool B. RNU 46 and RNU 48 were used as endogenous controls for data normalization. Another control not related to human is also included as a negative control. Each TaqMan Assay was run in quadruplicate. RNU 46 and RNU 48 expression was consistent in all the samples and displayed good range of CTs values 22–24 ) where as in ‘No Template’ Control (NTC) CT value was above 38. The average CT values of total profiled miRNAs in all samples were normalized with RNU 46 & RNU 48 using spotfire (statminer) software and fold change were represented in terms of log 10, 2 - Δ Δ CT (RQ ) and log10RQ. Only valid and significant miRNAs were picked up for further analysis.

ORGANISM(S): Homo sapiens

PROVIDER: GSE60386 | GEO | 2014/08/14

SECONDARY ACCESSION(S): PRJNA258157

REPOSITORIES: GEO

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