Home Technique / Molecular Biology / RNA transcriptional and post-transcriptional regulation

DNase I hypersensitivity assay (1)

Northern blot hybridization (3)

Nuclear run-on assay (1)

Primer extension (6)

RNA extraction and purification (5)

RNase protection assay (9)

S1 nuclease assay (6)


RNA methods from scienceboard.net new protocol
Protocol from Scientific Advisory Board (scienceboard.net). content 10/17/2003 RNA from tissues. 10/17/2003 Total cellular RNA 10/17/2003 EXTRACTION AND PURIFICATION OF TOTAL RNA USING TRIzol 10/19/2003 Native Gel Electrophoresis of R ...

mRNA purification protocol new protocol
mRNA purification protocol from Breeden's lab. The mRNA purification steps include preparing oligo-dT cellulose column, total RNA elution and washing in the oligo-dT cellulose column, precipitation and recovering of mRNA. Buffer recipes for mRNA purification a ...

Differential display of mRNA analysis (Breeden's lab) protocol
Differential display of mRNA analysis (Breeden's lab). Protocol link ...

Methods to Remove DNA Contamination from RNA Samples new protocol
An ambion tech notes on how to remove DNA contamination from RNA preparations. Genomic DNA false positive signals are easily identified by performing a "no-RT" control during RT-PCR. We can therefore assess the effectiveness of DNA removal methods by agaros ...

In Vitro Transcription Reverse Transcription new protocol
PDF prootcol by Paul Kroeger from Marimoto lab at Northwestern University. ...

Nonsense-mediated mRNA decay (NMD) new recommended review
Nonsense-mediated mRNA decay (NMD) is a mRNA degradation mechanism that eliminates mRNA containing premature translation-termination codons (PTC) usually at 55bp upstream of the exon junction complex (EJC). The NMD process is tightly coupled with translation a ...

Ten Ways to Improve Your RNA Isolation review
A tech notes from Ambion. It gives suggestions on following ten aspects: 1. Use proper cell or tissue storage conditions. 2. Thoroughly homogenize samples. 3. Pretreat homogenate before RNA isolation. 4.Choose the best RNA isolation method. 5. DNase tr ...

List of human house keeping gene new review
In this short review authors summarized use of human house keeping gene in gene expression study. Human housekeeping genes are compact. Eisenberg E, Levanon EY Trends Genet. 2003 Jul ; 19(7): 362-5 This web page lists all popular house keeping genes b ...

TechNote on Working with RNA new review
A technique bulletin describing the general guidance on working with RNA (Ambion Inc.). - Living with RNase. - RNA storage. - RNA precipitate - RNA radioactivity incorporation and yield. ...

Plasmid DNA extraction from e.coli for digestion mapping clony screening new review
Plasmid DNA extraction from e.coli for digestion mapping clony screening: This protocol utilize the Qiagen buffer P1, P2 and P3 from Qiagen plasmid purification kit. RNase must be added before use. It is cost efficient since it doesn't use expensive plasm ...

Redissolving RNA Pellet troubleshooting
I am working with total RNA from Chondrocyte cells. I am isolating the RNA using TRizol Reagent and I am having a problem with the last step of redissolving the RNA pellet. These cells have been put in 1.2% alginate and cultured for 5 days and then ex ...

mRNA isolation kits troubleshooting
I want to isolate mRNA from human B-Lymphocyte cell lines. Could somebody suggest some mRNA isolation kits which has given good yields. I will be using the mRNA for RT-PCR. ... ... ...

DNA contamination in RNA sample troubleshooting
I'm trying to look for a efficient method to remove the DNA contamination in my total RNA samples. I'm working with a Gram-positive bacteria E. hirae and I'm trying to use the RNA for quantitative RT-PCR. I've used both the Gaunidium thiocyanate method an ...

how to make nuclease free water ? troubleshooting
could anybody provide me with a short protocol on how to prepare nuclease free water? I need it for PCR, RT-PCR and RNA extraction from plant tissues. We have a milli-Q system and I don't want to use DEPC. ... ... ...

total RNA measurement troubleshooting
what would be a best choice of RNA standard (origin, size, etc., maybe even catalog #?) when comparing total RNA content in different species (both pro- and eucaryotes). ... ... ...

RNA extraction troubleshooting
I have isolated total RNA from non adherent cells with the TRIZOL reagent. At the end of the procedure, I solubilized the pellet with DEPC-treated water. After this, I measured OD 260 and 280 and calculated the ratio 260/280. This ratio should be 1.6 ...

Small scale TRIZOL extraction troubleshooting
I'd very much appreciate if somebody lets me know a recommended volume of TRIZOL for small size samples (10-20ul). Click here for replies to this question ...

General reverse transcription troubleshooting guide new troubleshooting
Low cDNA yield: 1. Check reverse transcription temperature. Different reverse transcriptase may require different reaction temperature, ranging from 37c to 65c. 2. RNA template is degraded or concentration is wrong. Use RNase inhibitor in the reac ...

RNA analysis tools and programs new software
This is a collection of RNA analysis tools from Pasteur Institute BioWeb. MFOLD: prediction of RNA Secondary Structure by Energy Minimization (Zuker) (advanced form) Vienna RNA package (documentation) RNAfold: calculate secondary structures of RNAs (adv ...

Antisense technonology animation media
Flash antisense technonology animation.(University of Nebraska - Lincoln) ...

More RNA protocols

RNA analysis
()
RNA can be purified in either total RNA or poly(A)-selected RNA (mRNA) form. mRNA comprises less than 5% of cellular total RNA. Generally speaking, for most RNA related applications, total RNA is sufficient for most requirements. Two specific applications, cDNA library construction and cDNA labeling for microarray require isolation of mRNA. Around 30ug of mRNA equels 1mg of total RNA.

Northern blot hybridization is used for determination of mRNA molecular weight and relative abundance. It is commonly performed using total RNA. Dot hybridization can not determine the size of mRNA species but can assay the quatity of the RNA target. Ribonuclease protection assay is also used to assy the abundance of mRNA species. RT-PCR is the most sensitive method for quantitatively measureing of mRNA expression level.

hnRNA isolation
(Forum)
protocoll for isolation of nuclear RNA


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