Do I need to make a special plasmid for bisulfite methylation test?

I have an inducible expression of the enzyme in a newly derived bacterial strain, and methylation may be affected. I want to test methylation efficiency with a bisulfite kit. The kit is the EZ DNA...

06 January 2021 7,218 2 View

How to perform cell transfection and Dual Glo assay with low-adhesive (HEK293) cells?

Hello everyone. I need to use Promega Dual Glo assay for HEK293 cells, but I am really afraid of wash out my cells from the 96-well plate, and at the same time, I am afraid to use coating because...

09 September 2020 4,817 7 View

How to reduce auto fluorescence in bone-marrow smears that would be used for FISH and IHC?

We are processing clinical samples of bone marrow smears, and have encountered autofluorescence that makes subsequent FISH and IHC virtually impossible. The problems are: 1. the samples are of...

18 December 2019 4,676 4 View

Antibody detects protein which was ablated in null mice (global knockout). Anyone had similar experience?

Here is a dilemma: there is a line of null mice (global knockout), which are null by genotyping and by mRNA analysis (qPCR). They were generated by replacement of an entire protein-coding region...

16 March 2019 9,722 6 View

Does high fidelity taq polymerase get less efficient after expiration date?

24 September 2016 5,468 4 View

Wrong or no fragment insert in directional TOPO cloning?

17 July 2016 10,020 10 View

Why in the analysis of cyanide, ninhydrin dissolved in sodium carbonate?

I was doing research related to the analysis of cyanide with ninhydrin, in some journals mentioned that preparation ninhydrin by dissolving it in sodium carbonate, but I do not understand why. Is...

12 July 2016 8,541 4 View

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15 June 2016 5,924 6 View

Why do I get a wrong length of PCR result but after sequencing it is actually a right target sequence?

Can somebody help me? I am doing a PCR of worm tissue. I used KAPA Taq Extra HotStart ReadyMix. I should be getting a PCR result of 158bp but I always get a 200 bp PCR result and after I sequence...

15 June 2016 7,313 6 View

What are the causes of PCR failure? i change the PCR master mix, optimized the annealing temperature and so on. but still no defined product band?

i want to amplified Channa striata COI gene using universal fish primer, out of 13 samples only two showed defined product bands after several optimization. I change the PCR mix, change the...

06 March 2016 854 6 View

Should we change media after1st 6hrs after plating HUVEC bought from INVITROGEN?it's not recomended by invitrogen.Will DMSO present in stock matter?

We have HUVEC bought from Invitrogen. Cat no -C-003-5c. while first plating the cells we have bought the protocol suggests not to centrifuge the cell with media to remove the DMSO. Should we...

02 March 2021 3,713 2 View

Diltuions for a Western blot?

I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...

22 February 2021 9,901 3 View

What are alternatives for B27+ Supplement in neuronal cell culture?

Hello everybody In our iPSC-derived neuronal culture we use Maturationmedium, supplemented with B27+. As Thermo Fisher run out of stock and can deliver earliest in May, I wanted to ask whether...

16 February 2021 3,348 1 View

How to revive a Hyphomicrobium strain?

11 February 2021 3,524 2 View

Bradford assay for insulin: discrepancies between standard curve and sample concentration during crystallisation?

Hi there, I would like to determine the unknown concentration of human insulin in my sample during crystallisation (the concentration range is roughly 0.5 -2 mg/ml, whereas the initial...

06 February 2021 9,370 7 View

How to measure volume of Pinus roxburghii ?

I want to measure volume of Pinus roxburghii in the Jammu region of India to calculate the carbon stocks of Pinus roxburghii.

31 January 2021 2,311 3 View

Where can I find the information about steel final production in Japan?

I am doing research about the in-use stock of iron and steel in Japan. I need the report about annual yield of steel final production. I found some information in the webpage of Japan Iron and...

25 January 2021 3,867 2 View

How to revive bacteria from glycerol stock?

19 January 2021 7,025 4 View

How can I prepare the erythromycin stock?

I need to prepare a stock of 10 mg / ml erythromycin to check the mic value. I am trying to dissolve 0.1g/10 ml of water but i can't dissolve erythromycin in water. How can i prepare 10 mg/ml...

03 January 2021 9,947 3 View

Which form of tryptophan and tyrosine is highly soluble in water?

The solubility of tryptophan and tyrosine in water is low. Is there a very highly soluble structurally similar or derivatives of these amino acids I can use? it doesn't matter if it is...

20 December 2020 3,157 3 View