[[Category:RNA]]
* Purified RNA
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1 KB (154 words) - 15:16, 24 July 2019
*Find the refseq RNA sequence for your gene by going to [http://www.ncbi.nlm.nih.gov/gene Entrez
...NA molecule. Pick 2-3 sequences, based on the alternative splicing of the RNA. If there is only one listed splice variant on NCBI Gene, pick one towards
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1 KB (173 words) - 22:37, 8 February 2012
*Add T7 RNA Polymerase sites to the 5' of each primer ('''TAATACGACTCACTATAGG''')
===RNA Synthesis===
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2 KB (290 words) - 14:31, 1 October 2009
*Calculate RNA to transfect (typically 40 pmol per well)
*Lipofectamine(in uL) = RNA(in nmoles) * 50
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1 KB (148 words) - 16:39, 18 December 2009
* Purify total RNA, via the protocol: [[ Purification of miRNA and mRNA with TRIzol ]]
===PolyA Tailing of RNA===
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3 KB (432 words) - 15:24, 1 March 2017
===RNA Extraction===
#Use RNEasy kit with Qiashredder. see [[Harvesting RNA from Cells grown in monolayer]] or [[Preparation_of_RNA_Samples_from_Mouse_
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3 KB (423 words) - 00:42, 29 March 2011
*PureLink RNA Mini Kit (Invitrogen cat#12183-018A)
...roform phase, an interphase and a colorless upper phase which contains the RNA.
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2 KB (355 words) - 17:07, 25 October 2017
*PureLink RNA Mini Kit (Invitrogen cat#12183-018A)
...pper phase which contains the RNA. (WAT tissue may separate into 3 phases, RNA is the top, colorless phase.
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3 KB (478 words) - 20:51, 22 October 2018
* You will then prepare a tube for each primer/RNA set as such in a PCR tube:
** 5 uL total RNA (presumes this is 1-10 ng)
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1 KB (234 words) - 18:52, 30 December 2014
# first 5 and last 5 are mX (methylated RNA nucleotides)
* notes: * = phosphorothiated; mX = 2' O-methyl RNA bases
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2 KB (373 words) - 20:43, 2 August 2011
...the spin colujmn membrane, ensuring that no ethanol is carried over during RNA elution. Residual ethanol may interfere with downstream reactions.
...gently, and centrifuge for 1 min at >8000 x g or >10,000 rpm to elute the RNA.
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3 KB (444 words) - 15:34, 12 May 2009
This was suggested by Invitrogen Technical Support, since the PureLink RNA prep kit is likely to lose much of the miRNAs. Ideally the PureLink mRNA o
# Centrifuge 15 min at 12 000g (11 000 RPM) at 4C to precipitate the RNA.
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2 KB (280 words) - 21:35, 29 December 2014
This protocol is for extracting DNA from TRIZOL preparations after the RNA-containing upper phase has been completely removed. It is based on the onli
* Perform TRIZOL extraction as you normally would for RNA extraction, ie by mechanically disrupting tissues in Qialyser. Approximate
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3 KB (417 words) - 17:42, 25 September 2015
[[ Category: RNA ]]
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687 bytes (92 words) - 18:59, 29 January 2017
* Total RNA, 1-5 ug
| RNA || 1-5 ug (from nanodrop)
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3 KB (551 words) - 21:04, 4 December 2012
#Find your RNA sequence on entrez by starting with the gene at http://www.ncbi.nlm.nih.gov
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1 KB (249 words) - 17:48, 29 June 2017
...3:''' A 6-well plate can also be used to increase surface area and protein/RNA content from each treatment. Seeding density of 90% followed by 2 days of g
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3 KB (473 words) - 20:02, 23 December 2013
*[[Harvesting RNA from Cells grown in monolayer]]
*[[Preparation of RNA Samples from Mouse Tissues]]
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4 KB (484 words) - 12:10, 15 August 2016
[[ Category: RNA ]]
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2 KB (306 words) - 16:49, 4 December 2018
[[ Category: RNA ]]
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2 KB (334 words) - 19:17, 18 December 2018