[IDT] PrimeTime qPCR Assay PCR Protocol -1

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ÀÌ protocolÀº Primetime qPCR Assay¸¦ »ç¿ëÇÏ´Â »ç¶÷µéÀ» À§ÇÑ °ÍÀÔ´Ï´Ù. ³óµµ¿Í ºÎÇÇ´Â ´Ù¸¥ product¿¡ ÀÇÇØ ´Þ¶óÁú ¼ö ÀÖ½À´Ï´Ù.

 

PrimeTime qPCR Assay Resuspension Protocol

 

1.     ¿î¹ÝµÇ´Â µ¿¾È ¹°ÁúµéÀÌ ºÐ¸®µÈ ¶§´Â Á¦Ç°À» ¿­±â Àü¿¡ PrimeTime qPCR Tube¸¦ 750Xg, 10ÃÊ µ¿¾È centrifuge¸¦ µ¹¸³´Ï´Ù.

2.     IDTE(100mM Tris, 0.1mM EDTA pH8.0)buffer¿¡¼­ÀÇ resuspensionÀÇ ¿ë·®Àº ¾Æ·¡ÀÇ Â÷Æ®¸¦ º¸½Ã¸é µË´Ï´Ù.

a.     PrimeTime qPCR Assay´Â 40X, 20X ¶Ç´Â 10X ³óµµ¿¡¼­ resuspensionÀÌ °¡´ÉÇÕ´Ï´Ù. ÀÌ·¯ÇÑ ¿©·¯ ³óµµ´Â ¾Æ·¡ÀÇ Â÷Æ®¿Í °°Àº final reaction volume ±×¸®°í, AssayÀÇ size¿¡ ÀÇÇØ ´Þ¶óÁý´Ï´Ù.

b.     °¡Àå È¿°úÀûÀÎ °á°ú¸¦ ¾ò±â À§ÇØ ¼¯¾îÁÝ´Ï´Ù.

3.     ResuspensionµÈ Assay¸¦ 750Xg, 10ÃÊ µ¿¾È centrifuge¸¦ µ¹¸³´Ï´Ù. ResuspensionµÈ Assay´Â ¾Æ·¡ÀÇ ¼³Á¤ Á¶°ÇÀÏ ¶§, ÃÖÁ¾ Yield°ªÀº 1XÀÇ 500nM Primers, 250nM Probe°¡ µË´Ï´Ù.

 

Recommeded Resuspension Volume for PrimeTime Assay Stock Creation

 

Final Desired Stock Concentration

40X

20X

10X

PrimeTime Mini qPCR Assay

Not recommeded

100¥ìl

200¥ìl

PrimeTime Std

qPCR Assay

250¥ìl

500¥ìl

1,000¥ìl

PrimeTime XL

qPCR Assay

1,250¥ìl

Not recommeded

Not recommeded


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