Detailed Steps on Primer Design

I list here the steps on how to design a good working primer.

1.       Get the sequence of your gene  from published literature for example OsNAS3.

2.       Using the TIGR Blast/ NCBI Blast, check the whole sequence if it specifies the gene of interest. It is important that the gene sequence is 100% homologous to the plant where the sequence is derived and the most important is that the whole sequence specifies only one distinct gene.

3.       Put your sequence inside the box of   Primer 3 online software. Specify in the software what are the parameters of the primer you want to have. Click generate. You would get something like this.

Important Notes:

When designing primers take note of the following:

GC content, melting temp (Tm) of primer, 3’ ends of the primer

PCR product size- It depends on what your purpose is. If your going to used the conventional PCR you can create a primer with a product size of 400bp and up. If you’re going to use the primer for real time PCR design a primers with 100-350 size only.  

4.       To be extra sure if your primer is really  specific,  Blast them again with NCBI blast. This would look like this. With the use of Vector NTI/Bioedit you can check the size of your PCR product.

5.       You are now ready to order your primers at SBS and proligo.

Erythrocyte Sedimentation Rate

Erythrocyte sedimentation rate (ESR) AKA  sedimentation rate or Westergren ESR, is the rate at which red blood cells sediment/precipitate in a duartion of one hour. It is a very common blood test, and is a NON-specific measure of inflammation. In order to conduct this test, anticoagulated blood is placed in an upright tube, known as a Westergren tube, and the rate at which the red blood cells fall is measured and reported in mm/h. 

Diagnostic Use: 
 ESR increased can be triggered  by any cause or focus of inflammation.It may be increased in pregnancy, inflammation, anemia or rheumatoid arthritis. However it may  be  decreased in polycythemia, sickle cell anemia, hereditary spherocytosis, and congestive heart failure. Studies reiterated that it may be increased in kidney cancer.It should be taken note that basal ESR is slightly higher in females(Medline).

Commonly Use:
 useful in diagnosing some diseases such as:
 multiple myeloma, 
temporal arteritis,
 polymyalgia rheumatica,
 systemic lupus erythematosus, 
rheumatoid arthritis,
chronic kidney diseases.

 In many of such cases, the ESR may exceed 100 mm/hour.

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