figure2b: miR34a asRNA doxorubicin treatment of HCT116 p53 wt and null.

In GranderLab/miR34a_asRNA_project: The miR34AasRNAproject package facilitates the transparency and reproducibility of the miR34a asRNA project and associated publication.

Description

HCT116 cells were cultured in DMEM high modified (Hyclone, GE healthcare) supplemented with supplemented with 2mM L-glutamine, 50ug/ml Penicillin-Streptomycin and 10 were plated in 6 well plates. 24 hours later media was exchanged and doxorubicin was added to a final concentration of 100, 200 or 500 ng/ml. Cells were harvested for RNA extraction 24 hours later using trypsin. RNA was extracted using Nucleospin RNA kit (Machery-Nagel Ref. 740955) according to manufacturer<e2><80><9f>s protocol and DNase treated using Ambion Turbo DNA-free according to manufacturer<e2><80><9f>s protocol (Life Technologies Ref. AM1907). cDNA was synthesized using ~500 ng RNA with M-MLV (Life Technologies Ref 28025013) and a mixture of oligo(dT) with nanomers in accordance with the manufacturer's protocol. qPCR quantification was carried out using the PowerUp SYBR Green Master Mix (Thermo Fisher Scientific, Ref. A25777) on the CFX96 Touch Real-Time PCR Detection System: 50<c2><b0>C for 2 min, 95<c2><b0>C for 2min, and 95<c2><b0>C for 1 sec followed by 60<c2><b0>C for 30 sec repeated for 40 cycles.

Format

A tibble with 72 rows and 7 variables:

'Cell line'

Character; Indicating the cell line the data corresponds to.

Condition

Factor; Indicating the genetic status of TP53 in the cell line.

Treatment

Factor; Indicates the treatment (doxorubicin) concentration (ng/ml).

'Biological Replicate'

Numeric; Indicates the biological replicate of the sample.

gene

Factor; indicates the gene that the Ct values correspond to.

Ct1

Numeric; Ct value corresponding to the first technical replicate.

Ct2

Numeric; Ct value corresponding to the 2nd technical replicate.

...

Examples

getData('Figure 2b')

GranderLab/miR34a_asRNA_project documentation built on May 26, 2019, 7:26 a.m.