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Questions related to Molecular Biological Techniques
Hi, I had set up PCR with 50ng of FFPE and i was able see band at expected size. when i tried to repeat the same PCR with 50ng each in 4 vials i:e to bulk up the product. i did not get any band...
12 January 2017 3,056 1 View
In general from a confluent flask first I trypsinize and pellet down the cells, re suspend in 2 ml of media.Then I take 100ul of cell suspension and add 4 volume of trypan blue.After 5 mins, the...
11 January 2017 6,825 4 View
Hello Everyone, I am trying to develop a protein expression and purification system. Hence, can anyone tell me about "Intein Tags" or related papers, articles or suggestion? Please Help Thanks
09 January 2017 3,171 3 View
Hi all! While doing my DNA isolation I encountered a new problem. After the addition of chloroform and during centrifugation, the ependorff and 50mL centrifuge tubes melted. Can anyone provide me...
09 January 2017 2,766 12 View
Details: IMAGE ATTACHED Load 50ug protein in 20ul (All samples denatured for 10mins at 75% Precast 12% thermo gels Run 90volts 15mins (stack) 150volts for remainder of gel. Also run on...
09 January 2017 4,075 9 View
I am going to express Human CD36 on a mammalian cells, so I wonder if there's a company that I could buy the construct in a plasmid, and if they also provide positive and negative controls. Regards
04 January 2017 8,107 2 View
I am looking for rough recommended concentrations but I have only found in vitro treatment concentrations.
02 December 2016 6,415 1 View
I am currently trying to prepare a library for illumine deep sequening using the yeast Candida glabrata. Ive been having difficulty generating a high quality library. My collaborators believe that...
28 November 2016 1,047 1 View
i have 3 groups, negative control, injected normal cells, injected mutation cells. when i got the counts/sec, i found the control group is also high, do i need to use cells group to divide NC...
24 November 2016 1,560 1 View
Is there anyone who ever used dual-crosslinking method in ChIP experiment? I want to ask if there are any subsequent effect on the Chromatin fragmentation step, and because increase the spacer...
22 November 2016 4,812 3 View
Hi, can anybody send me an optimized protocol for short fragment of sscp pcr (117bp) using sliver staining?
22 November 2016 7,602 3 View
We are trying to isolate CD63+ exosomes in order to characterize them by electronic microscopy, Nanosight and ultimately by proteomics/transcriptomics. To isolate them we use the “Total Exosome...
21 November 2016 1,175 16 View
There are so many microbial diversity investigations using high-throughput sequencing on variety of natural environments, e.g. land soil, fresh and salt water, sediment etc. Some of them sampling...
21 November 2016 4,983 4 View
I cut pcmv6-AC-ha-his(origene) with hind111 and xho1, the same as my insert whose size is about 1k. Then I ligate them which labeled as "target" ,and negative control is the ligation system which...
15 November 2016 7,176 3 View
I'm trying to isolate RNA from suspended adipocyte now. The problem is that after centrifuging, the cells were still floating at the bottom, so when I remove all the media before adding RLT...
14 November 2016 2,276 4 View
I did DNA extraction of cyanobacteria. Up until the very end it seemed to be working fine. The Chloroform/isoamly alcohol (24:1) steps all formed a separate layer when introduced and seemed to be...
09 November 2016 7,254 6 View
I want to detect gene expression by multiplex PCR. The multiplex PCR reaction works fine on genomic DNA, but on a cDNA template the same oligo mix seems not to work as nicely, although specific...
07 November 2016 8,661 5 View
I just did my first RNA extraction from tissue using a trizol-like reagent called "Tranzol". I must have eluted it to 70ul final volume but I'm having poor yields of approximately 150ng/ul, low...
31 October 2016 6,561 5 View
Recently, i'm doing a sonication to bacterial pellet using PBS 10x buffer. I'm being confused in adding the buffer to the pellet for sonication. I'm setting the amplitude for 20-30%. Is there any...
28 October 2016 8,997 2 View
How to make Tris Succinate buffer (0.05M, pH8.2)?
26 October 2016 5,153 3 View
I tred to dissolve vinclozolin for cell culture use. I dissolved it in DMSO. There was no problem in this step. For further dilution, I was used full medium (DMEMF12+FBS), and water, seperatly....
25 October 2016 3,806 1 View
I want to constructed Crispr Cas system in archeae and I need to know which promoter and terminater I should use to express RNAguide. Indeed, if I understand well, promoter should be one for tRNA...
18 October 2016 1,346 3 View
Hi. How can I remove endotoxin from bacteriophage lysate with a simple and high-efficiency method?I will appreciate to help me.
18 October 2016 4,645 1 View
Is amino benzoxymethyl paper the best choice?
16 October 2016 2,013 3 View