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02 March 2019 2,656 1 View

Why the cDNA not coming good from a good RNA?

03 April 2016 9,393 13 View

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02 March 2016 7,893 10 View

How can I increase the yield of longer capped transcripts for in vitro transcription?

I am setting up in vitro reaction by T7 polymerase on linearized plasmids purified by Promega column after digestion. Some people recommend using phenol;chloroform to clean DNA before...

09 October 2015 2,564 6 View

Is it possible to silence a transgene (driven by a strong promoter) by an artificial miRNA driven by an inducible promoter in Arabidopsis?

While in literature there is no report, to my knowledge, to silence one transgene in Arabidopsis (under 35S promoter) using another transgene (artificial microRNA, under inducible system). I think...

05 June 2015 2,416 1 View

Is the trizol method to isolate RNA from plant tissues good for RNA-seq experiments?

We normally isolate RNA from rice and Arabidopsis by using trizol. We would like to do RNA-seq and small-RNA seq. There are commercial kits like miRVana which allow to isolate total RNA including...

02 March 2015 624 5 View

While doing VIGS, can the infiltrated virus spread from the leaves to the germline tissues and knockdown the germline gene?

We have seen VIGS induced silencing of PDS, GFP and many other genes in many species. Is there any report or is it possible that we can knockdown a gene expressed in germline tissues using VIGS?...

31 December 2014 7,572 3 View

Which is the best kit to isolate plant total RNA (inclucing miRNA)?

I want to isolate total RNA (including siRNA and miRNA) from Arabidopsis seedlings. We want to do RNA-seq and small RNA-seq with that RNA. There are few kits available for this purpose to isolate...

08 September 2014 1,436 14 View

What is the right hygromycin concentration to select rice (Nipponbare) calli?

Hi, I am selecting rice calli of Nipponbare on hygromycin (20mg/L) on selection as well as regeneration media. Some protocols suggest more than 30mg/L and even 50. What is the optimal...

08 September 2014 8,116 15 View

Is pTF101.1 binary vector good to transform Arabidopsis?

08 September 2014 4,219 4 View

Chlorinated organic formation in odor control scrubbers using hypochlorite?

I'm involved in a study of odor control technologies for municipal wastewater treatment plant. One of the control options involves a chemical 2-stage (acid/alkaline) packed bed scrubber. The...

03 March 2021 3,661 2 View

What solvent can I used for Paal Knorr reaction between 6-amino hexanoic acid and 2.5 hexanedione?

I am try to make the Paal Knorr reaction between 2.5 hexnedione and 6-amino hexanoic acid, my problem is type of solvent to use, because 6-hexaminohexanoic acid is soluble in water but I am not...

02 March 2021 4,443 2 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

Ester cleavage conditions?

Hi all, I need to cleave the ester in Ethyl 2-amino-1H-imidazole-5-carboxylate to make the carboxylic acid and retrieve the product from solution to use in downstream applications of linking to...

01 March 2021 8,766 3 View

How to remove DCU and HOBT from my reaction?

I am doing a acid and a chiral amino alcohol reaction using DCC and HOBT in DCM or THF? Can anyone suggest me how to remove DCU and HOBT from a reaction mixture??

28 February 2021 2,153 1 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

What is the most common number of epitopes in an multi-epitope Vaccine?

hi everyone, I'm interested in the field of vaccine design specifically multi-epitope cancer vaccines , I want to know if there is a minimum and maximum number of epitopes we could use in a...

27 February 2021 3,882 3 View

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 View

How to increase solubility of Poly acrylic acid in Dioxane solvent?

I am working with Polyacrylic acid (100.000Da) and instead of water, I need to use another solvent (dioxane) to dissolve PAA. However, because of the high molecular weight, it's quite hard to...

25 February 2021 2,225 5 View