RNAsimple Total RNA Kit

RNA isolation / purification Cells - immortalized H1299

Experiment
RNA isolation / purification Cells - immortalized H1299
Product
RNAsimple Total RNA Kit from Tiangen
Manufacturer
Tiangen

Protocol tips

Upstream tips
- To wipe off
RNase, the glassware can be roasted at 150℃ for 4 hours,
while plastic can be dipped in 0.5 M NaOH for 10min, washed
by RNase-Free ddH2O thoroughly, and sterilized

Publication protocol

Total RNA in each sample was extracted by an RNA extraction kit (Tiangen Biotech Co., Ltd., Beijing, China) completely following the manufacturer instructions and reverse-transcribed into cDNA. RT-qPCR was carried out by Stratagene Mx3000P (Stratagene; Agilent Technologies, Inc., Santa Clara, CA, USA) using Bestar® SybrGreen qPCR MasterMix (DBI® Bioscience, Ludwigshafen, Germany) with the following protocol: Initial denaturation at 95°C for 2 min, 40 cycles consisting of 94°C for 20 sec, 58°C for 20 sec, and 72°C for 20 sec. The following primers were used: miR-126-5p, 5′-CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGCGCGTA-3′ (sense) and 5′-ACACTCCAGCTGGGCATTATTACTTTTGGTA-3′ (antisense); VEGFR-A, 5′-AGGGCAGAATCATCACGAAGT-3′ (sense) and 5′-AGGGTCTCGATTGGAT-GGCA-3′ (antisense); VEGFR-2, 5′-ATAGAAGGTGCCCAGGAAAAG-3′ (sense) and 5′-GTCTTCAGTTCCCCTCCATTG-3′ (antisense); MEK1, 5′-GGGCTTCTA-TGGTGCGTTCTA-3′ (sense) and 5′-CCCACGGGAGTTGACTAGGAT-3′ (antisense); ERK, 5′-TCTGGAGCAGTATTACGACCC-3′ (sense) and 5′-CTGGCTGGAATCTAGCAGTCT-3′ (antisense); β-actin, 5′-ATCGTGCGTGACA-TTAAGGAGAAG-3′ (sense) and 5′-AGGAAGGAAGGCTGGAAGAGTG-3′ (antisense). Relative mRNA expressions were obtained by 2−∆∆CT method. β-actin was served as an internal control.

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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?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA isolation / purification Cells - immortalized H1299 using RNAsimple Total RNA Kit from Tiangen.

Paper title
Antitumor effects of Tubeimoside-1 in NCI-H1299 cells are mediated by microRNA-126-5p-induced inactivation of VEGF-A/VEGFR-2/ERK signaling pathway
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Manufacturer protocol

Download the product protocol from Tiangen for RNAsimple Total RNA Kit below.

Download PDF Download manufacturer protocol

Videos

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