Dewi Yuliana

15 Questions 34 Answers 0 Followers

Questions related from Dewi Yuliana

How long I can storage cells after fixation with paralformaldehide (PFA)? Is okey for long term?

Dear all, I want to try filipin staining using MEF cell culture. I have planned for not used immediately after fixation. How long can I storage the cell after fixation with paraformaldehyde...

02 February 2017 9,006 0 View

Why I got smear in my PCR product using kod plus?

Dear all, I have done PCR by using KOD FX with two step method ( 5 cycle with Tm 50 and 35 cycle with Tm 55, and elongation time 1.30 minute with size 1356bp, then I got single band without...

11 November 2016 6,539 5 View

How to separate 404 bp with 344 bp for dna product PCR?

Dear All, My target gene have another variant and it was expressed too in my gel. my target gene is 404 bp and the variant 344. how to longer separate it. so I can cut my target gene and...

09 September 2016 1,268 3 View

What the reason rasio vector : dna is 1: 90 by using vector p.BluecriptSK-II-ecoRV?

Dear all, , I want to know what the best basic theory we need use rasio 1:90 between vector : dna with using p.BluecriptSK-II digested by EcoRV? best regards, Dewi Yuliana

07 July 2016 983 7 View

How to design phosphorylation PCR product ?

Dear ALL I want to phosphorylate my PCR product, please give me sugestion how to calculate p mol PCR product.., is there any protocol without converse.., is there any link or literature I can use...

07 July 2016 5,710 1 View

Is no problem if I do Ethanol precipitation with composition glicogen, Na-asetat, before and after phosphorylation PCR product by T4 PNK ?

Dear all, Is there anyone have experience do ethanol precipitation PCR product prior to phosphorylation by T4 polynucleotide kinase? According literature ethanol precipitation by ammonium asetat...

07 July 2016 3,231 3 View

Can anyone help with running gel for purification by gel extraction?

Dear All I have done gel extraction my PCR product (2300bp), but the final I lose all my DNA. Here my following step. Ethanol precipitation (using glycogen, Na-OCH, ethanol) My PCR product 50...

07 July 2016 4,280 4 View

Is it possible to Clone ArfGAP protein with SH3 domains, ankyrin repearts & pleckstrin homology domains (ASAP1) in neuron brain mouse tissue?

Dear All I have plan to clone ASAP1 from neuron brain mouse tissue. Is there anyone have experience cloning ASAP1 with cDNA from neuron brain mouse? I already design primer and check by primer...

06 June 2016 247 0 View

The meaning any, self, pair any and pair End by using primer3plus ??

Dear all I have already design Forward Primer and Reserve primer Primer F = Tm 69,8C GC 73,7% Any 6,0 and Self 3,0 Primer R = Tm 69,8C GC 54,0% Any 3,0 and Self 0 Pair any ; 5 and pair end...

05 May 2016 9,447 3 View

How to solve Unexpected band after amplification PCR ?

I have done amplification by PCR for my gene, but unexpected I got several band higher expression than my band, I have try changed my condition annealing time (Tm) but the result still same,...

05 May 2016 7,318 20 View

How to check cDNA mouse embryonic fibroblast gene expression ?

Dear All Is there any software to know gene expression high or low in mouse embryonic fibroblast? Best Regards

04 April 2016 1,151 12 View

How to avoid cells aggregation in mcf7 before measurement by FACS to get single clone?

Dear all, I have tried knock out cells in mcf7 cells then shorting, by FACS, but I got problem, My cells always got aggregation, so I can't got single clone. Please give me any suggestions or...

12 December 2015 6,764 5 View

How do I grow single cell become colonies after sorting by FACS in mcf7 cells?

I have already prepared knock out cell by using px459 for knock out mcf7 cell then shorting by using FACS to get single clone, one day after FACS I get around 10 single clone/well ( positive EGFP...

12 December 2015 8,176 7 View

Is there a possibility that the virus accepted by the cells after several days was rejected from the cells, in a stable transfection of a cell line?

I had injected retrovirus containing pBabe.puro.cmv and pcl ampho to the F1 B16 and then after 24 hours and 48 hours I did puromycin selection, The First days and second days the cell decrease...

04 April 2015 8,241 13 View

I had done miniprep pEGFP-N1 but I get low band in my miniprep and I get 2 band in running p-EGFP-N1?

Its the first time I work with plasmid pEGFP-N1 but when I runned the gel agarose get two band and my band mini preparation is low. Please give me suggestion why I get two band? thanks very...

04 April 2015 2,373 11 View