ISOGEN

RNA isolation / purification Cells - immortalized CHO

Experiment
RNA isolation / purification Cells - immortalized CHO
Product
ISOGEN from Nippon Gene
Manufacturer
Nippon Gene

Protocol tips

Upstream tips
- This kit can be used for the isolation of RNA from up to 1x10^7 cells

Publication protocol

Total RNA was isolated from CHO or UPS-1 cells using Isogen (Nippon Gene) or RNeasy Mini Kit (Qiagen) and then subjected to RT-PCR analysis using the SuperScript III One-Step RT-PCR system (Invitrogen). For quantitative RT-PCR (qRT-PCR), total RNA was subjected to reverse transcription using a SuperScript VILO cDNA Synthesis Kit (Invitrogen). The resultant cDNA was used as a template for PCR using LightCycler FastStart DNA MasterPLUS SYBR Green I (Roche Applied Science); fold changes in gene expression were normalized to the β-actin mRNA level. Chinese hamster ATP11C cDNA was amplified using the following primer pairs: set 1 (sense, 5′-ATACTGAGCTCTTAGAACTGACC-3′; antisense, 5′-ATCA­CTATTCCTGGCTGCTTGG-3′), set 2 (sense, 5′-GAACAGCACA­TCA­ACGTTGATAC-3′; antisense, 5′-CTGATAAATATGAGGAGAATTATGG-3′), and set 3 (3′ untranslated region (UTR); sense, 5′-GTATAGGGTTCAGAATAAATGTCC-3′; antisense, 5′-GATATTAGACCAAGACAATTAGTC-3′). The ATP11A, CDC50A, and β-actin cDNAs were amplified using the following primer pairs: sense, 5′-CATGGAAGTGCTCAA­GAGAGAC-3′/antisense, 5′-GAGCAGGCTGACAGTGACAA­G-3′; sense, 5′-GCCA­GTTAAATGGAGACCCTAG-3′/antisense, 5′-GTCC­AGCTGGTAATGTTGGATG-3′; and sense, 5′-CTGTATGCCTCTGGTCGTAC-3′/antisense, 5′-GCCATCTCCTGCTCGA­AGTC-3′, respectively.

Full paper   Login or join for free to view the full paper.

Reviews

ISOGEN from Nippon Gene has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Discussion

5 years ago

Author: Switzerland

I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?

I am having trouble using the QiageN RNAeasy mini kit for CD34+ cells. Any tips?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Cells - immortalized CHO using ISOGEN from Nippon Gene.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Nippon Gene for ISOGEN below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing RNA isolation / purification Cells - immortalized CHO using ISOGEN from Nippon Gene. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms