25 Questions 12 Answers 0 Followers
Questions related from Lalita Sharma
Hello everyone, I am looking for the protocol to calculate MIC and MBC by broth microdilution method but I am unable to get a suitable protocol. In papers, they have given the reference of CLSI...
18 March 2021 5,311 7 View
Hello everyone, I have to isolate DNA from Brucella phage , but before that, I have to concentrate the phage using PEG 8000. I have got protocol for concentrating the phage using PEG and TM buffer...
25 January 2021 3,087 6 View
Hello everyone, In eukaryotic cells, all the mRNAS are polyadenylated at 3' end. I want to ask whether this is the same case for pox viruses also. Do all their mRNAs are polyadenylated at 3'end...
04 May 2019 9,540 2 View
Dear all, I have a DNA sample with 5 ug/ul conc. but it's 260/280 ratio is 1.21 and 260/230 ratio is 1.55 which indicates that DNA is not pure and has some impurities. So, I want to know whether...
26 March 2019 4,821 4 View
Hello everyone I have bisulphite converted my DNA using kit from Zymo research now I have to do methylation specific PCR. I have to check effect of my drug on methylation status of diff. genes...
16 November 2017 277 4 View
Hello everyone, I isolated DNA from freshly taken Human blood but during processing I was getting RBC contamination in pellet .I washed with RBC lysis buffer 6-7 times followed by...
17 October 2017 7,758 3 View
Hello everyone? I have done HDAC activity assay using kit from epigenetics. But my standard curve didnot come properly. Has anyone used the same kit? If used then please send the standard graph...
01 June 2017 8,084 0 View
I have done methylation specific PCR for p21 and p27 with prostate cancer cell lines, LNCaP and DU145 but I didnot get any band for methylated as well as unmethylated allele for both the genes. I...
26 February 2017 6,054 6 View
I have done bisulphite treatment manualy as well as with kit from zymo and then methylation specific PCR. but , I am getting primer dimer in 2 out of 3 genes and no bands in 2 other genes in both...
09 November 2016 6,882 6 View
Hello, I want to do radioactivity based DNMT activity assay. Can somebody upload the whole prtocol for this? Thank you
28 July 2016 9,364 5 View
I have done RT -PCR for Cyclin D2, p21 and p27 in du145 , but did not get any band . in p21 , there was v. faint band but when I repeated the same , I did not get any band for p27. does these 3...
14 April 2016 4,646 1 View
hello, I am working on rccs. I started with 12 rpm speed for 5 ml and 10 ml vessel in RCCs for glial cells ( doubling time:24 hrs) and seeding density of 2*105, but while harvesting I can see...
29 March 2016 2,702 0 View
we purchased RCCS from synthecon Inc. In users guide it is mentioned to prepare vessels by rinsing with media or PBS for 5 mins to overnight. I want to know the purpose of doing this...
24 January 2016 2,790 2 View
I am doing MSP after BS treatment.. I standarized the protocol with different conc. of BS but when I did with my samples, I have faced many problems? I have 6 samples but first time I got in only...
04 October 2015 1,022 1 View
I want to do simple RT- PCR for some genes. so, can I use the primer seq. which are used for real time PCR for these genes. The product size of real time PCR is generaly v. small. so if the...
16 September 2015 3,690 3 View
I want to check the global methylation status in some cancer cell line. I want to know whether I can use direct sequencing of PCR product of repetitive elements after Bisulphite treatment for...
19 August 2015 4,085 3 View
I have extracted DNA from a cell line. I usually get 5 ug DNA each time when I resuspend in 20 ul water. But this time the max. DNA I obtained was 2ug with same cells, same protocol, same time...
08 July 2015 2,214 6 View
I have methylated my DNA of interest using M. Sss1 methyl transferase from NEB in 20 ul reaction set up as mentioned in their protocol.After this, I tried to purify that DNA by phenol extraction...
29 May 2015 6,515 3 View
We have Two types of DU145 cells in our lab and I am badly confused which one to use. So, Please upload some photos of DU145 so that I can come out of the confusion. Thank you
18 December 2014 1,350 3 View
I have some solution contining beta mercaptoethanol. I want to check its pH. somebody told me not to use pH meter for this. so plz suggest I can use H meter or not
24 November 2014 9,240 1 View
I want to use LNCaP for some molecular studies.So,I just wanted to know the highest passage number of this cell line which can be used for these kind of studies
08 September 2014 4,979 7 View
I am getting a non specific band in PCR. So, I want to do hot start PCR. I want to know while doing hot start PCR manually, we have to add polymerase after initial denaturation step of 94 degree...
01 August 2014 4,407 8 View
I want to induce cancer in c.elegans using some chemical. so can somebody please tell me the chemicals which can be used for this purpose? Is it possible with benzopyrene, DMBA, diethylnitrosamine...
16 July 2014 4,522 1 View
Is anybody currently doing bisulphite treatment or methylation specific PCR? I am doing bisulfite treatment and then methylation specific PCR, but not getting the desired result.So, I want to know...
01 July 2014 5,946 6 View
I am doing methylation specific PCR for various genes. I did bisulphite treatment referring protocol by Herman et al. In my methylation specific PCR result for all the genes ,I am not getting...
25 June 2014 7,777 3 View
