Designing forward primer for mi cDNA?

I am using Agilent miRNA first strand synthesis kit (cat#600036) for preparing mi cDNA from total RNA. A universal primer is provided with the kit sequence of which is not given. i want to study...

07 August 2018 2,227 2 View

No signal in SYBR Green QPCR for miRNA quantification?

Hello everyone ! I am using mirVana miRNA isolation kit for the isolation of total RNA containing small RNA. Further i used Agilent first strand synthesis kit(cat#600036) for the synthesis of...

07 August 2018 750 5 View

Problem with PCR amplification?

i tried gradient PCR for a new set of primers and got the amplification. However, when i tried running the same set of primers on a single annealing temperature, it showed no or weak...

10 November 2016 9,245 14 View

Can anyone tell how to troubleshoot no amplification in PCR?

I am trying to amplify my genes of interest.Though I got the amplified product of the desired size on my first attempt , i am not able to get the amplification again. What might be the problem and...

07 August 2016 9,433 3 View

Problem with RNA Isolation using mirVAna kit?

I have been using mirVana miRNA isolation kit for total rna isolation and had a good experience using that. Recently, i am facing problem with the kit. 280/230 ratio and concentrations are too low...

03 April 2016 5,013 3 View

Problem with RNA isolation from Breast tissue ?

02 March 2016 1,431 3 View

Why do I get peaks in NTC in qPCR ?

I am performing real time pcr for ACTB. I am getting a ct value of 17-20 in my samples. However , i am getting a peak in NTC too with a ct value around 33-35. Also the dissociation curve for NTC...

10 November 2015 3,707 8 View

What factors should be considered in miRNA -target prediction?

06 July 2015 8,356 6 View

Can anyone help explain double bands in PCR?

06 July 2015 4,069 6 View

How can I select a good endogenous control for miRNA qPCR?

I have to perform qPCR for miRNA isolated from colorectal tumor and adjacent normal tissue samples. Is U6 snRNA a good endogenous control for qPCR?

05 June 2015 3,646 9 View

In PRIMER, is it possible to downweight certain taxa in multivariate community datasets to reflect uncertainty around identification??

I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...

03 March 2021 8,066 4 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

How to calculate BREQ 3 questionnaire?

how to calculate BEHAVIOURAL REGULATION IN EXERCISE QUESTIONNAIRE BREQ 3 ,which contain 24 questions and each question has 5 answers (0- Strongly disagree , 1-disagree, 2- Neither agree nor...

02 March 2021 811 3 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

I am interested in medicinal chemistry field, which books are highly recommended to learn from?

I am at the beginner level in drug design and drug discovery, and I am interested to learn more about this field. Which books are highly recommended to start from and gives me all essential...

02 March 2021 8,437 2 View

Please suggest a good text book for "Research Methodology in Life Sciences". Which book would be good for PhD Scholars?

Please provide Book Title and author name

02 March 2021 9,059 3 View

Could any one guide me, to write research proposal for grant? ?

We have one plant, the local people using this for controlling blood sugar, the toxicity and antidiabetic activity of the plant is not reported yet, please guide from whag should be the process ?...

02 March 2021 2,300 1 View

How to calculate the annealing temperature of a primer pair?

I have to amplify a gene and my primers just reached. The Tm for Forward primer is 64.2, and that of reverse primer is 65.5. Can some one suggest how to get the best annealing temperature? Thanks...

01 March 2021 360 7 View

Anybody with an idea on how do design specific primers for detecting tomato resistance genes tm-1, TM-2 and Tm2^2?

I am trying to identify these 3 genes among some tomato cultivar collections and after aligning some sequences from NCBI, I couldn't find unique sequences to target for specific primers. There...

28 February 2021 606 3 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