杭州泰禾生物技术有限公司-杭州泰禾生物技术有限公司

PNA 合成

PNA 产品

多肽合成

PNA Clamp Kit

技术介绍 PNAClamp EGFR Kit PNAClamp KRAS Kit PNAClamp PIK3CA Kit 订购产品

PNAClamp 技术是指PNA能够专一性的与野生型DNA结合，从而使野生型DNA不能作为模板进行PCR扩增；而突变型DNA，由于碱基的缺失或突变，PNA不能与其结合，从而使突变型DNA能够进行PCR扩增。

一般来说，一个样本中，大部分是野生型DNA，突变型DNA只占极少的一部分，因此通过一般的PCR很难检测到突变型DNA的存在。如果在样本中加入与野生型DNA互补的PNA，则野生型DNA与PNA牢固结合，使得野生型DNA不能进行PCR扩增，由于没有野生型DNA的干扰，突变性DNA能够很容易被扩增并检测。

PNAClamp Mutation detection 的原理：

1. 与 DNA/DNA 的结合相比，在不存在错配的情况下，PNA/DNA 的结合稳定性更强（每增加 1 base 序列，退火温度增加～1℃）。

2. 与 DNA/DNA 的结合相比，在存在错配的情况下，PNA/DNA 的结合稳定性更差。

3. PNA 不能作为引物被 DNA polymerase 识别并扩增。

PNAClamp Mutation detection Kits 的优势：

1. 检测时间短（约2小时）。

2. 操作简单。

3. 需要样品少（低至1～25ng DNA样品）。

4. 灵敏度高。

5. 稳定性强。

6. 保质期长。

7. 可以在多种型号的荧光定量PCR仪中使用（如：BI-7500, ABI-7900HT, Biorad CFX-96/384, Roche Lightcycler, Qiagen RGQ etc）。

8. 数据分析简单明了。

PNAClamp Mutation detection 示意图：

The epidermal growth factor receptor (EGFR) is a family member of Receptor tyrosine kinases, expressed on the surface of epidermal cells. Overexpression or overactivation of EGFR is linked to a number of cancers, including lung cancer, anal cancers and glioblastoma multiform.

The PNAClamp EGFR Mutation Detection Kit detects most prevalent mutations described to date in the EGFR gene, including T790M, the presence of which correlates with resistance to tyrosine kinase inhibitors. Detecting somatic mutations in EGFR gene may provide a useful strategy to predict the response to the tyrosine kinase inhibitors in efforts to increase the survival rate of lung cancer patients receiving targeted therapy.

Exon	Amino acid change	Base change
18	Gly719Ala	2156 G>C
	Gly719Ser	2155 G>A
	Gly719Cys	2155 G>T
19	Glu746_Ala750del	2235_2249 del 15
	Glu746_Thr751delinslle	2235_2252 AAT (complex)
	Glu746_Ser752del	2236_2253 del 18
	Glu746_Thr751delinsAla	2237_2251 del 15
	E746_S752>A	2237_2254 del 18
	Glu746_Ser752delinsVal	2237_2255 >T (complex)
	Glu746_Ala750del	2236_2250 del 15
	Glu746_Ser752delinsAsp	2238_2255 del 18
	L747_A750>P	2238_2248 >GC (complex)
	Leu747_Thr751delinsGln	2238_2252 >GCA (complex)
	Leu747_Glu749del	2239_2247 del 9
	Leu747_Thr751del	2239_2253 del 15
	Leu747_Ser752del	2239_2256 del 18
	Leu747_Glu749del:Ala750Pro	2239_2248 TTAAGAGAAG>C
	Leu747_Pro753delinsGln	2239_2258 >CA (complex)
	Leu747_Thr751delinsSer	2240_2251 del 12
	Leu747_Pro753delinsSer	2240_2257 del 18
	Leu747_Thr751del	2240_2254 del 15
	Leu747_Thr751deinsPro	2239_2251 >C (complex)
20	Thr790Met	2369 C>T
	Ser768lle	2303 G>T
	Ala767_Val769dupAlaSerVal	2307_2308 ins9
	His773dupHis	2319_2320 insCAC
	Asp770_A771insGly	2310_2311 insGGT
21	leu858Arg	2573 T>G
21	leu861Gln	2582 T>A

KRAS mutation is found in several cancers including colorectal, lung, thyroid, and pancreatic cancers and cholangiocarcinoma. KRAS mutations are often located within codons 12 and 13 of exon 2, which may lead to abnormal growth signaling by the p21-ras protein. These alterations in cell growth and division may trigger cancer development as signaling is excessive.

A KRAS mutation often serves as a useful prognostic marker of drug response. For example, a KRAS mutation is considered to be a strong prognostic marker of response to tyrosine kinase inhibitors such as gefitinib (Iressa) or erlotinib (Tarceva). Recently, KRAS mutations have been detected in many colorectal cancer patients and may be associated with responses to cetuximab (Erbitux) or panitumumab (Vectibix), which are used in colon cancer therapy.

Codon	Mutation	Base change
Codon12	Gly12Asp	35G>A
	Gly12Ala	35G>C
	Gly12Val	35G>T
	Gly12Ser	34G>A
	Gly12Arg	34G>C
	Gly12Cys	34G>T
Codon13	Gly13Ser	34G>T
	Gly13Arg	37G>C
	Gly13Cys	37G>T
	Gly13Asp	38G>A
	Gly13Ala	38G>C
	Gly13Val	38G>T
Codon59	Ala59Ser	175G>T
	Ala59Thr	175GA
	Ala59Glu	176C>A
	Ala59Gly	176C>G
	Ala59del	176_178 del CAG
Codon61	Gly60Asp	179G>A
	Gly60Ala	179G>C
	Gly60Val	179G>T
	Gly60Gly	180T>A
	Gly60Gly	180T>C
	Gln61Glu	181C>G
	Gln61Lys	181C>A
	Gln61Leu	182A>T
	Gln61Arg	182A>G
	Gln61Pro	182A>C
	Gln61His	183A>T
	Gln61His	183A>C

Codon 117	Lys117Glu	349A>G
	Lys117Arg	350A>G
	Lys117Asn	351A>C
	Lys117Asn	351A>T

Codon 146	Ala146Pro	436G>C
	Ala146Thr	436G>A
	Ala146Gly	437C>G
	Ala146Val	437C>T
	Ala146Ala	438A>G
	Ala146Ala	438A>C
	Ala146Ala	438A>T

PI3K (Phosphoinositide 3-kinases or PI 3-Kinases) are family of lipid kinases capable of phosphorylating the 3' position hydroxyl group of the inositol ring of phosphatidylinositol. They are involved in coordinating a diverse range of cell functions including cell growth, proliferation, differentiation, motility, survival and intracellular trafficking.

Activating mutations in PI3K catalytic domain of the p110alpha subunit (PIK3CA) have recently been discovered in certain types of cancer cells. PIK3CA mutations are found at 25~40 % frequency in various types of tumors including colorectal cancer, gastric cancer, lung cancer, brain cancer, endometrial cancer, ovarian cancer, breast cancer. About 80% of the point mutations presides in exon 9 (a presumed helical domain) and exon 20 (a presumed kinase domain), while other types of mutations are also seen in different locations.

Tube No	Reagent	Amino Acid Change	Base change	Exon	Translation region
1	E542	Glu542Lys	1624 G>A	Exon 9	Helical
		Glu542Gly	1624 A>G
		Glu542Val	1624 A>T
2	E545	Glu545Lys	1633 G>A
		Glu545Gly	1634 A>G
		Glu545Asp	1635 G>T
3	H1047	His1047Tyr	3139 C>T	Exon 20	Kinase
		His1047Leu	3140 A>T
		His1047Arg	3140 A>G

产品信息：

Catalog No.	Product Name	Description	Size	Price
PNAC-1002	PNAClamp KRAS Kit (v2)	G12, G13	30 tests	Inquire
PNAC-1003	PNAClamp KRAS Kit (v3)	G12, G13, Q61	25 tests	Inquire
PNAC-1004	PNAClamp KRAS Kit (plus)	A59, K117, A146	25 tests	Inquire
PNAC-1006	PNAClamp KRAS Kit (v4)	G12, G13, A59, Q61, K117, A146	25 tests	Inquire
PNAC-1101	PNAClamp NRAS Kit (v4)	G12, G13, A59, Q61, K117, A146	25 tests	Inquire
PNAC-2001	PNAClamp BRAF Kit	BRAF V600 mutation	50 tests	Inquire
PNAC-3002	PNAClamp EGFR Kit	G619, E19 del, T790, S768, E20 in, L858, L861	25 tests	Inquire
PNAC-4002	PNAClamp PIK3CA Kit	E542, E545, H1047	25 tests	Inquire
PNAC-5001	PNAClamp IDH1 Kit	R132 mutation	25 tests	Inquire