27 Questions 25 Answers 0 Followers
Questions related from Lalita Sharma
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...
05 May 2019 3,288 2 View
Hello everyone. I want to do DNMT activity analysis by using prostate cancer cells. I want to know whether we can store the nuclear lysate for 2-3 days in -20 degree before performing activity...
06 June 2018 6,537 7 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...
11 November 2017 8,356 4 View
I am going to do ms pcr but I have doubt on water milliQ water because previously RT pcr result was not coming by using that water. So I borrowed water from another department but that water was...
10 October 2017 3,988 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...
10 October 2017 5,423 3 View
Hello, I have 3- 4 month old DNA stored at - 2O degree. Can I use thisDna for bisulphite treatment followed by methylation specific pcr? Thank you
01 January 2017 5,347 5 View
Hello, I want to do radioactivity based DNMT activity assay. Can somebody upload the whole prtocol for this? Thank you
07 July 2016 2,232 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...
04 April 2016 906 1 View
What is the best way to freeze LNCaP cells? ATCC recommends complete medium plus 5% DMSO. some people use 10% DMSO, 20% FBS and rest complete medium. Some use 90% FBS and 10% DMSO. Please tell me...
01 January 2016 2,427 3 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...
01 January 2016 4,465 2 View
I have to use LNCaP cells for Methylation specific PCR, but LNcaP cells are not available but LNCaP FGC cells are available. So Can I use these cells in place of LNCaP cells?
11 November 2015 6,592 3 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...
10 October 2015 589 2 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...
09 September 2015 5,410 4 View
I have 2 month old lysate. I want to do western blot analysis for HDAc 1-7 using this lysate. Can I use this lysate? In other words I want to ask about protein stability of HDAcs in the lysate,...
04 April 2015 4,732 7 View
I am having a problem with identification of DU 145 cells. There are 3 types of DU cells in our lab. 2 are quiet similar but the third one is v. different. I am uploading the pics of all three 10X...
01 January 2015 1,449 9 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
12 December 2014 8,402 4 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
11 November 2014 4,417 2 View
I am doing western. I prepared sample buffer using .5 M TRIS which was diluted from 2.5 M TRIS buffer stock having ph 6.8, I did not check the ph of .5 M TRIS and made the sample buffer using this...
11 November 2014 2,306 8 View
I want to do methylation specific PCR. So, I am doing bisulfite treatment. In some protocols 3M stock of sodium bisulfite is used which finally becomes 2.6 M in solution (Herman et al). some uses...
10 October 2014 9,525 9 View
I did Western blotting for HDAC1 (1:1000 dilution of antibody from cell signaling). I got v. faint band after 1 hour exposure. I stripped the membrane and increased the dilution to 1:200 and this...
10 October 2014 6,768 16 View
I am maintaining LNCaP cells and facing various problems. I was observing black dots in my culture previously.so, I re -filtered media and started growing fresh cells. That problem ended there ,...
09 September 2014 8,761 21 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
09 September 2014 4,721 8 View
I am maintaining the LNCaP cells. I am observing black dots in background medium. Initially, the dots were less in no. but they increased after subsequent passages and now my cells are having...
08 August 2014 7,296 16 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...
08 August 2014 3,118 9 View
I isolated DNA from one of the prostate cancer cell line, its 260/280 ratio is 1.95 and 260/ 230 ratio is 1.31. After DNA isolation, I did Bisulphite treatment and after this treatment 260/280...
07 July 2014 386 10 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...
07 July 2014 1,833 1 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...
06 June 2014 5,821 3 View
