FlashTag™ Biotin HSR RNA Labeling Kits

Microarray RNA amplification & Labeling - Human blood Biotin

Experiment
Microarray RNA amplification & Labeling - Human blood Biotin
Product
FlashTag™ Biotin HSR RNA Labeling Kits from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Upstream tips
Always keep thawed reagents and reaction tubes on ice

Publication protocol

For each sample, 400 ng of total RNA was labelled with the FlashTag Biotin RNA labelling kit for Affymetrix GeneChip miRNA 2.0 arrays (Affymetrix, Santa Clara, CA, USA) according to the manufacturer’s instructions. A tailing reaction was carried out at 37°C for 15 min in a volume of 15 µL reaction mix (10 µL of total RNA and spike control oligos mix, 1 × reaction buffer, 1.5 mM MnCl2, ATP, and 1 mL poly A polymerase enzyme) followed by the ligation of the biotinylated signal molecule to the target RNA sample at 25°C for 30 min (with the addition of 4 µL of 5 × FlashTag Ligation Mix Biotin and 2 µL of T4 DNA Ligase into the 23.5 mL of reaction mix). Finally, 2.5 mL of stop solution was added to stop the reaction. After labelling, the samples were hybridised on Affymetrix GeneChip miRNA 2.0 arrays (Affymetrix, Santa Clara, CA, USA) at 48°C and 60 rpm for 16 h in a volume of 103.2-µL reaction mix (which contained 21.5 µL of total biotin-labelled RNA, 50 µL hybridisation mix, 15 µL formamide, 10 µL DMSO, 5 µL eukaryotic hybridisation controls, and 1.7 µL control oligonucleotide B2). Immediately after hybridisation the arrays were washed and stained with streptavidin phycoerythrin conjugate to remove nonspecific binding. Finally, they were scanned to determine miRNA expression levels.

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

Reviews

FlashTag™ Biotin HSR RNA Labeling Kits from Thermo Fisher Scientific has not yet been reviewed for this experiment

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

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Microarray RNA amplification & Labeling - Human blood Biotin using FlashTag™ Biotin HSR RNA Labeling Kits from Thermo Fisher Scientific.

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

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for FlashTag™ Biotin HSR RNA Labeling Kits below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Microarray RNA amplification & Labeling - Human blood Biotin using FlashTag™ Biotin HSR RNA Labeling Kits from Thermo Fisher Scientific. 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.

Become shareholder Discussions About us Contact Privacy Terms