x********u
发帖数: 430 | 1 The procedure I found in the literature;
1) grow E. coli to late-exponential phase
2) extract total RNA with RNeasy Protect Bacteria Mini Kit(Qiagen)
3) purify RNA by RNeasy Mini Kit(Qigen) spin column
4) send samples to xxxxx University Functional Genomics Core Facility for
amplication, labeling and hybridization to the Affymetrix GeneChip E. coli
genome 2.0 Array.
5) Data analysis by MatLab bioinformatics toolbox
Question: For amplication of total mRNA and labeling of the resulting cDNA
in step 4, does they have a generic primer which can reverse transcribe the
whole population of the mRNA? Thanks. |
f*******e
发帖数: 628 | 2 是 mRNA 的话，一般都利用 polyA tail 来 reverse transcription. |
x********u
发帖数: 430 | 3 If I am not wrong, I remember there is No polyA tail for prokaryotic cell
like E coli
【在 f*******e 的大作中提到】
: 是 mRNA 的话，一般都利用 polyA tail 来 reverse transcription.
|
f*******e
发帖数: 628 | 4 那就用 random primer， 一般的 kit 都会自带那种 6-mer 的 primer.
【在 x********u 的大作中提到】
: If I am not wrong, I remember there is No polyA tail for prokaryotic cell
: like E coli
|
