07 July 2016 1 6K Report

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 ?

Best Regards

More Dewi Yuliana's questions See All
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...

01 February 2017 9,027 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...

10 November 2016 6,569 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...

08 September 2016 1,294 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

06 July 2016 1,004 7 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...

06 July 2016 4,306 4 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...

06 July 2016 3,258 3 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...

05 June 2016 272 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...

04 May 2016 9,494 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,...

04 May 2016 7,354 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

03 April 2016 1,170 12 View

Similar questions and discussions
How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 View

How to analyze multiple phosphorilation sites?

Hello, I am currently analyzing some phosphoproteomics data, but I have peptides with multiple phosphorylation sites or phosphorylations together with carbamidomethylation or oxidation. How can I...

04 August 2024 8,432 3 View

Does anyone have issues using Prepman Ultra reagent for MicroSeq ID bacterial, fungal and yeast sample preparation?

I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...

01 August 2024 2,079 0 View

PCR showing no bands - Master mix and primers didn't mix?

Hello everyone, I performed a PCR yesterday, and the results showed no bands on the gel. Of course, I probably missed some crucial steps, like adding my samples to the PCR strips themselves, for...

31 July 2024 2,406 6 View

Dimensions of an MJ Research 96-well alpha module?

Can anyone with an MJ research / BioRad PCR machine from ~2010 or earlier tell me the external measurements (LxWxH) of the removable standard PCR alpha modules that can be removed from a PTC200 or...

30 July 2024 2,867 0 View

How to retain amplicon yield after Ampure bead cleanup, following a 2-stage PCR protocol?

Hello all, I have been trying to follow a 2-stage PCR protocol used to amplify barcodes of a large yeast library, as per Nyugen et al. (2022) -...

30 July 2024 841 2 View

Someone have the key or the installer of the program Mx pro 3000 (PCR real time)?

I need to install this program

25 July 2024 4,756 0 View

Illustra™ MicroSpin™ G-25 columns what it is used for?

Hello, We found three packages of Illustra™ MicroSpin™ G-25 columns in the cabinet of an unused lab. They are very old but have never been opened. I have never used this kit before, and I couldn't...

25 July 2024 4,927 3 View

Our partners will collect data and use cookies for ad personalization and measurement. Learn how we and our ad partner Google, collect and use data. Agree & close