Harshraj Subhash Shinde

13 Questions 41 Answers 0 Followers

Questions related from Harshraj Subhash Shinde

Is it possible that Crisper Cas9 will cut the DNA at downstream of PAM site?

I did crisper cas9 experiment in plants, after creating the transgenic plants i analyzed the sequence for digestion and i saw digestion of single base at downstream of PAM site (5 base pair after...

09 September 2017 4,967 6 View

Which is the most common method of promoter isolation in plant?

I want to isolate the promoter of my gene for promoter analysis. so i need to isolate the full length sequence of promoter. So can anyone tell me the most commonly used method for isolation of...

11 November 2016 549 11 View

Can anyone suggest basic protocol for 5' and 3' RACE PCR without Kit?

Doing 5' and 3' RACE with kit is bit expensive . can anyone share protocol of 5' and 3' RACE PCR without Kit or some literature regarding. I am trying to amplify unknown 3' and 5' region of my...

06 June 2016 2,250 4 View

Is it possible to perform the PCR using oligo dt primer and Gene specific primer using RNA as a template?

I want to know weather is it possible to perform the PCR using oligo dt primer and Gene specific primer using RNA as a template? if possible then how to calculate the annealing temperature to set...

05 May 2016 1,987 3 View

Can anyone help me to read the GFP subcellular localization images?

To determine the exact subcellular localization of my gene within plant cell. GFP gene was fused in frame with my gene. Then i performed transient expression analysis by introducing fusion gene...

05 May 2016 654 7 View

Can anyone suggest me Rapid and Highly efficient method of tobacco transformation?

To study the function of my gene. I have cloned my gene in PBi vector and then i have moved the vector to Agrobacterium tumefaciens strain EHA 105. now I am thinking to proceed for tobacco...

04 April 2016 345 6 View

Where to buy k667 yeast mutant strain?

To study the calcium and sodium transport function of one transporter gene I want to study them in k667 yeast mutant strain. So can anyone guide me me how and where to buy this strain? Thank you...

03 March 2016 5,726 5 View

How to isolate full length of gene ?

I did Multiple Sequence Alignment of three genes and then I designed degenerate primers using the homologous (conserved) region. After PCR of C-DNA in another species using degenerate primer i got...

02 February 2016 9,651 13 View

Why am I having problems with the isolation of a 1.4 Kb gene from rice cDNA?

I am trying to isolate a gene from rice whose size is 1.4 Kb. I have synthesized cDNA. Then In PCR of C-DNA I am getting amplification in House keeping gene (actin) but in case of my gene of...

01 January 2016 7,196 17 View

How does a difference in GC content in primers affect PCR?

I am trying to amplify C-DNA by RT-PCR for that I using forward primer with GC content 68.5 and reverse primer with GC content 53. After performing PCR I am not able to amplify my gene. So this...

12 December 2015 4,040 11 View

What is quality distribution in RNA sequencing data?

What is distribution of average sequence quality scores in RNA seq data analysis?

10 October 2015 9,302 2 View

How to read the dots of scatter plot while studying the gene expression level?

Does any one knows how do read the dots of scatter plot while studying the gene expression level? Please guide by checking the attached file as a reference. Thank you.

10 October 2015 1,269 4 View

Can anyone tell me how to grow natural accessions of Arabidopsis on 1/2 MS media in GWAS?

Recently I have started working on GWAS. I got Arabidopsis natural accessions from ABRC to study the phenotypes. But I don't know how to grow these plant. Weather on just 1/2 MS media or on media...

10 October 2015 2,352 3 View