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Questions related from Manita Raina
I have been doing qpcr for an year now but everytime I am getting errors. Either positive control cq values will vary or there will be no amolification. The knockdown genes are showing increased...
08 June 2024 7,165 1 View
I had mammalian cancer cell line cultured and fixed for confocal imaging. While observing I found these particles around cell nuclei stained with dapi. I checked the main culture for bacteria but...
16 May 2024 8,988 2 View
I had mammalian cancer cells cultuted and fixed on confocal slides. I could see these small small bodies stained with dapi. The main culture flask doesnt show any bacterial growth. Is this...
16 May 2024 6,008 3 View
I was running sds page 8% gel and it started running tilted(see the picture attached). Can anyone tell what can be the reason behind gel running like this?
18 January 2024 7,723 0 View
I revived 2 skov3 cell lines together in a t-25 flask. These were stored in -80 for more than 6 months. Their morphology is changed. They look stressed. But can anyone what can be the cause of stress?
23 December 2023 6,752 0 View
I am trying create a mutant plasmid using DpnI digestion. I had put pcr using the mutant primers. But then as I could not do immediate transformation, I kept it to store in -20 degree. Now if I...
09 September 2023 3,598 1 View
I am trying to develop my western blot using an antibody targeting a 17kda protein. It is binding non-specifically and not giving any band(like in the photo). What does it mean is happening? How...
23 August 2023 3,446 4 View
I tried to revive a2780 cells cryopreserved in -80 degrees. Its been more than 12 hours and its still not attaching. What could possibly be happening? What should I do?
22 August 2023 820 4 View
I designed primers which have the tm 69 is and the gc content is 41%. I tried increasing gc content upto 50% but then its increasing primer tm to 72. If I keep gc content 41%, will my primers...
30 June 2023 7,188 2 View
I am trying to transfect an sh-rna in my mammalian cells using lipofectamine 2000 reagent but every time transfection does not completely. Im using 1.5 ug of dna 5ul of lipofectamine and 200ul of...
13 June 2023 8,593 0 View
I have to clone a cDNA to insert in pcdna3.1. The forward primer generated is having tm 83 and reverse primer has tm 67. I am not getting amplification from normal pcr, gradient pcr, hotstart pcr...
06 June 2023 1,097 6 View
I need to amplify a gene cDNA using PCR reaction and need to check if it got amplified by Agarose gel electrophoresis method. My amplified cDNA should be around 3kb. What percent of Agarose gel...
26 February 2023 4,173 5 View
I need to design cloning pcr primers. I have made forward primer of 65 nucleotides and reverse primer of 32 nucleotides. Though the sequences from gene of interest for both primers are of same...
06 January 2023 3,972 3 View
